Compounds and methods for modulating GFAP

ABSTRACT

Provided are compounds, methods, and pharmaceutical compositions for reducing the amount or activity of GFAP RNA in a cell or subject, and in certain instances reducing the amount of GFAP in a cell or subject. Such compounds, methods, and pharmaceutical compositions are useful to ameliorate at least one symptom or hallmark of a leukodystrophy. Such symptoms and hallmarks include motor delays, cognitive delays, paroxysmal deterioration, seizures, vomiting, swallowing difficulties, ataxic gait, palatal myoclonus, autonomic dysfunction, and presence of intra-astrocytic inclusions called Rosenthal fibers. Such leukodystrophies include Alexander Disease.

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing is provided as a file entitledBIOL0353UWOSEQ_ST25.txt, created on Jul. 17, 2020, which is 592 KB insize. The information in the electronic format of the sequence listingis incorporated herein by reference in its entirety.

FIELD

Provided are compounds, methods, and pharmaceutical compositions forreducing the amount or activity of GFAP RNA in a cell or subject, and incertain instances reducing the amount of glial fibrillary acidic protein(GFAP) in a cell or subject. Such compounds, methods, and pharmaceuticalcompositions are useful to ameliorate at least one symptom or hallmarkof a leukodystrophy. Such symptoms and hallmarks include motor delays,cognitive delays, paroxysmal deterioration, seizures, vomiting,swallowing difficulties, ataxic gait, palatal myoclonus, autonomicdysfunction, and intra-astrocytic inclusions called Rosenthal fibers.Such leukodystrophies include Alexander Disease.

BACKGROUND

Alexander Disease (AxD) is a rare developmental disorder that affects˜1/1,000,000 live births and is caused by a number of differentautosomal dominant mutations in the gene encoding glial fibrillaryacidic protein, GFAP. AxD is a typically fatal leukodystrophy with earlyonset (<age 4, Type I) or later onset (>age 4 Type II) forms (Prust etal., (2011) GFAP mutations, age at onset, and clinical subtypes inAlexander disease. Neurol 77: 1287-1294). Symptoms include motor andcognitive delays, paroxysmal deterioration, seizures, encephalopathy,macrocephaly, and intra-astrocytic inclusions called Rosenthal fibers.

There are no specific therapies for AxD, with current treatments beinglimited to supportive treatments for individual symptoms (e.g.,antiepileptics to prevent seizures; Messing, et. al., “Strategies fortreatment in Alexander Disease”, Neurotherapeutics: The Journal of theAm. Soc. For Expt. NeuroTher., 2016).

Currently there is a lack of acceptable options for treatingleukodystrophies such as AxD. It is therefore an object herein toprovide compounds, methods, and pharmaceutical compositions for thetreatment of such diseases.

SUMMARY OF THE INVENTION

Provided herein are compounds, methods and pharmaceutical compositionsfor reducing the amount or activity of GFAP RNA, and in certainembodiments reducing the expression of glial fibrillary acidic proteinin a cell or subject. In certain embodiments, the subject has aleukodystrophy. In certain embodiments, the subject has AlexanderDisease (AxD). In certain embodiments, compounds useful for reducing theamount or activity of GFAP RNA are oligomeric compounds. In certainembodiments, compounds useful for reducing the amount or activity ofGFAP RNA are modified oligonucleotides. In certain embodiments,compounds useful for decreasing expression of glial fibrillary acidicprotein are oligomeric compounds. In certain embodiments, compoundsuseful for decreasing expression of glial fibrillary acidic protein aremodified oligonucleotides.

Also provided are methods useful for ameliorating at least one symptomor hallmark of a leukodystrophy. In certain embodiments, theleukodystrophy is Alexander Disease. In certain embodiments, the symptomor hallmark includes motor delays, cognitive delays, paroxysmaldeterioration, seizures, vomiting, swallowing difficulties, ataxic gait,palatal myoclonus, autonomic dysfunction, and presence ofintra-astrocytic inclusions called Rosenthal fibers.

DETAILED DESCRIPTION OF THE INVENTION

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive. Herein, the use of the singular includes theplural unless specifically stated otherwise. As used herein, the use of“or” means “and/or” unless stated otherwise. Furthermore, the use of theterm “including” as well as other forms, such as “includes” and“included”, is not limiting. Also, terms such as “element” or“component” encompass both elements and components comprising one unitand elements and components that comprise more than one subunit, unlessspecifically stated otherwise.

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.All documents, or portions of documents, cited in this application,including, but not limited to, patents, patent applications, articles,books, treatises, and GenBank, ENSEMBL, and NCBI reference sequencerecords, are hereby expressly incorporated-by-reference for the portionsof the document discussed herein, as well as in their entirety.

Definitions

Unless specific definitions are provided, the nomenclature used inconnection with, and the procedures and techniques of, analyticalchemistry, synthetic organic chemistry, and medicinal and pharmaceuticalchemistry described herein are those well-known and commonly used in theart. Where permitted, all patents, applications, published applicationsand other publications and other data referred to throughout in thedisclosure are incorporated by reference herein in their entirety.

Unless otherwise indicated, the following terms have the followingmeanings:

Definitions

As used herein, “2′-deoxynucleoside” means a nucleoside comprising a2′-H(H) deoxyfuranosyl sugar moiety. In certain embodiments, a2′-deoxynucleoside is a 2′-β-D-deoxynucleoside and comprises a2′-β-D-deoxyribosyl sugar moiety, which has the β-D ribosylconfiguration as found in naturally occurring deoxyribonucleic acids(DNA). In certain embodiments, a 2′-deoxynucleoside may comprise amodified nucleobase or may comprise an RNA nucleobase (uracil).

As used herein, “2′-MOE” or “2′-MOE sugar moiety” means a 2′-OCH₂CH₂OCH₃group in place of the 2′-OH group of a furanosyl sugar moiety. A “2′-MOEsugar moiety” means a sugar moiety with a 2′-OCH₂CH₂OCH₃ group in placeof the 2′-OH group of a furanosyl sugar moiety. Unless otherwiseindicated, a 2′-MOE sugar moiety is in the β-D-ribosyl configuration.“MOE” means O-methoxyethyl.

As used herein, “2′-MOE nucleoside” means a nucleoside comprising a2′-MOE sugar moiety.

As used herein, “2′-OMe” or “2′-O-methyl sugar moiety” means a 2′-OCH₃group in place of the 2′-OH group of a furanosyl sugar moiety. A“2′-O-methyl sugar moiety” or “2′-OMe sugar moiety” means a sugar moietywith a 2′-OCH₃ group in place of the 2′-OH group of a furanosyl sugarmoiety. Unless otherwise indicated, a 2′-OMe sugar moiety is in theβ-D-ribosyl configuration.

As used herein, “2′-OMe nucleoside” means a nucleoside comprising a2′-OMe sugar moiety.

As used herein, “2′-substituted nucleoside” means a nucleosidecomprising a 2′-substituted sugar moiety. As used herein,“2′-substituted” in reference to a sugar moiety means a sugar moietycomprising at least one 2′-substituent group other than H or OH.

As used herein, “5-methylcytosine” means a cytosine modified with amethyl group attached to the 5 position. A 5-methylcytosine is amodified nucleobase.

As used herein, “administering” means providing a pharmaceutical agentto a subject.

As used herein, “antisense activity” means any detectable and/ormeasurable change attributable to the hybridization of an antisensecompound to its target nucleic acid. In certain embodiments, antisenseactivity is a decrease in the amount or expression of a target nucleicacid or protein encoded by such target nucleic acid compared to targetnucleic acid levels or target protein levels in the absence of theantisense compound.

As used herein, “antisense compound” means an oligomeric compoundcapable of achieving at least one antisense activity.

As used herein, “ameliorate” in reference to a treatment meansimprovement in at least one symptom or hallmark relative to the samesymptom or hallmark in the absence of the treatment. In certainembodiments, amelioration is the reduction in the severity or frequencyof a symptom or the delayed onset or slowing of progression in theseverity or frequency of a symptom. In certain embodiments, the symptomor hallmark is motor delays, cognitive delays, paroxysmal deterioration,seizures, vomiting, swallowing difficulties, ataxic gait, palatalmyoclonus, autonomic dysfunction, and presence of intra-astrocyticinclusions called Rosenthal fibers.

As used herein, “bicyclic nucleoside” or “BNA” means a nucleosidecomprising a bicyclic sugar moiety.

As used herein, “bicyclic sugar” or “bicyclic sugar moiety” means amodified sugar moiety comprising two rings, wherein the second ring isformed via a bridge connecting two of the atoms in the first ringthereby forming a bicyclic structure. In certain embodiments, the firstring of the bicyclic sugar moiety is a furanosyl moiety. In certainembodiments, the furanosyl sugar moiety is a ribosyl moiety. In certainembodiments, the bicyclic sugar moiety does not comprise a furanosylmoiety.

As used herein, “cerebrospinal fluid” or “CSF” means the fluid fillingthe space around the brain and spinal cord. “Artificial cerebrospinalfluid” or “aCSF” means a prepared or manufactured fluid that has certainproperties of cerebrospinal fluid.

As used herein, “cleavable moiety” means a bond or group of atoms thatis cleaved under physiological conditions, for example, inside a cell,an animal, or a human.

As used herein, “complementary” in reference to an oligonucleotide meansthat at least 70% of the nucleobases of the oligonucleotide or one ormore portions thereof and the nucleobases of another nucleic acid or oneor more portions thereof are capable of hydrogen bonding with oneanother when the nucleobase sequence of the oligonucleotide and theother nucleic acid are aligned in opposing directions. As used herein,“complementary nucleobases” means nucleobases that are capable offorming hydrogen bonds with one another. Complementary nucleobase pairsinclude adenine (A) and thymine (T), adenine (A) and uracil (U),cytosine (C) and guanine (G), 5-methylcytosine (^(m)C) and guanine (G).Complementary oligonucleotides and/or target nucleic acids need not havenucleobase complementarity at each nucleoside. Rather, some mismatchesare tolerated. As used herein, “fully complementary” or “100%complementary” in reference to an oligonucleotide, or a portion thereof,means that the oligonucleotide, or portion thereof, is complementary toanother oligonucleotide or target nucleic acid at each nucleobase of theshorter of the two oligonucleotides, or at each nucleoside if theoligonucleotides are the same length.

As used herein, “conjugate group” means a group of atoms that isdirectly or indirectly attached to an oligonucleotide. Conjugate groupsinclude a conjugate moiety and a conjugate linker that attaches theconjugate moiety to the oligonucleotide.

As used herein, “conjugate linker” means a single bond or a group ofatoms comprising at least one bond that connects a conjugate moiety toan oligonucleotide.

As used herein, “conjugate moiety” means a group of atoms that isattached to an oligonucleotide via a conjugate linker.

As used herein, “contiguous” in the context of an oligonucleotide refersto nucleosides, nucleobases, sugar moieties, or internucleoside linkagesthat are immediately adjacent to each other. For example, “contiguousnucleobases” means nucleobases that are immediately adjacent to eachother in a sequence.

As used herein, “constrained ethyl” or “cEt” or “cEt modified sugarmoiety” means a 4′ to 2′ bridge in place of the 2′OH-group of a ribosylsugar moiety, wherein the bridge has the formula of 4

CH(CH₃)—O-2

and wherein the methyl group of the bridge is in the S configuration. A“cEt sugar moiety” is a bicyclic sugar moiety with a 4′ to 2′ bridge inplace of the 2′OH-group of a ribosyl sugar moiety, wherein the bridgehas the formula 4

CH(CH₃)—O-2

and wherein the methyl group of the bridge is in the S configuration.

As used herein, “cEt nucleoside” means a nucleoside comprising a cEtsugar moiety.

As used herein, “chirally enriched population” means a plurality ofmolecules of identical molecular formula, wherein the number orpercentage of molecules within the population that contain a particularstereochemical configuration at a particular chiral center is greaterthan the number or percentage of molecules expected to contain the sameparticular stereochemical configuration at the same particular chiralcenter within the population if the particular chiral center werestereorandom. Chirally enriched populations of molecules having multiplechiral centers within each molecule may contain one or more stereorandomchiral centers. In certain embodiments, the molecules are modifiedoligonucleotides. In certain embodiments, the molecules are compoundscomprising modified oligonucleotides.

As used herein, “chirally controlled” in reference to an internucleosidelinkage means chirality at that linkage is enriched for a particularstereochemical configuration.

As used herein, “deoxy region” means a region of 5-12 contiguousnucleotides, wherein at least 70% of the nucleosides are2′-β-D-deoxynucleosides. In certain embodiments, each nucleoside isselected from a 2′-β-D-deoxynucleoside, a bicyclic nucleoside, and a2′-substituted nucleoside. In certain embodiments, a deoxy regionsupports RNase H activity. In certain embodiments, a deoxy region is thegap or internal region of a gapmer.

As used herein, “gapmer” means a modified oligonucleotide comprising aninternal region having a plurality of nucleosides that support RNase Hcleavage positioned between external regions having one or morenucleosides, wherein the nucleosides comprising the internal region arechemically distinct from the nucleoside or nucleosides comprising theexternal regions. The internal region may be referred to as the “gap”and the external regions may be referred to as the “wings.” The internalregion is a deoxy region. The positions of the internal region or gaprefer to the order of the nucleosides of the internal region and arecounted starting from the 5′-end of the internal region. Unlessotherwise indicated, “gapmer” refers to a sugar motif. Unless otherwiseindicated, the sugar moiety of each nucleoside of the gap is a2′-β-D-deoxynucleoside. In certain embodiments, the gap comprises one2′-substituted nucleoside at position 1, 2, 3, 4, or 5 of the gap, andthe remainder of the nucleosides of the gap are 2′-β-D-deoxynucleosides.As used herein, the term “MOE gapmer” indicates a gapmer having a gapcomprising 2′-β-D-deoxynucleosides and wings comprising 2′-MOEnucleosides. As used herein, the term “mixed wing gapmer” indicates agapmer having wings comprising modified nucleosides comprising at leasttwo different sugar modifications. Unless otherwise indicated, a gapmermay comprise one or more modified internucleoside linkages and/ormodified nucleobases and such modifications do not necessarily followthe gapmer pattern of the sugar modifications.

As used herein, “hotspot region” is a range of nucleobases on a targetnucleic acid that is amenable to oligomeric compound-mediated reductionof the amount or activity of the target nucleic acid.

As used herein, “hybridization” means the pairing or annealing ofcomplementary oligonucleotides and/or nucleic acids. While not limitedto a particular mechanism, the most common mechanism of hybridizationinvolves hydrogen bonding, which may be Watson-Crick, Hoogsteen orreversed Hoogsteen hydrogen bonding, between complementary nucleobases.

As used herein, “internucleoside linkage” means the covalent linkagebetween contiguous nucleosides in an oligonucleotide. As used herein,“modified internucleoside linkage” means any internucleoside linkageother than a phosphodiester internucleoside linkage. “Phosphorothioateinternucleoside linkage or “PS internucleoside linkage” is a modifiedinternucleoside linkage in which one of the non-bridging oxygen atoms ofa phosphodiester internucleoside linkage is replaced with a sulfur atom.

As used herein, “leukodystrophy” means a disorder due to abnormalitiesin the myelin sheath of neurons.

As used herein, “linker-nucleoside” means a nucleoside that links,either directly or indirectly, an oligonucleotide to a conjugate moiety.Linker-nucleosides are located within the conjugate linker of anoligomeric compound. Linker-nucleosides are not considered part of theoligonucleotide portion of an oligomeric compound even if they arecontiguous with the oligonucleotide.

As used herein, “non-bicyclic modified sugar moiety” means a modifiedsugar moiety that comprises a modification, such as a substituent, thatdoes not form a bridge between two atoms of the sugar to form a secondring.

As used herein, “mismatch” or “non-complementary” means a nucleobase ofa first oligonucleotide that is not complementary with the correspondingnucleobase of a second oligonucleotide or target nucleic acid when thefirst and second oligonucleotide are aligned.

As used herein, “motif” means the pattern of unmodified and/or modifiedsugar moieties, nucleobases, and/or internucleoside linkages, in anoligonucleotide.

As used herein, “nucleobase” means an unmodified nucleobase or amodified nucleobase. As used herein an “unmodified nucleobase” isadenine (A), thymine (T), cytosine (C), uracil (U), or guanine (G). Asused herein, a “modified nucleobase” is a group of atoms other thanunmodified A, T, C, U, or G capable of pairing with at least oneunmodified nucleobase. A “5-methylcytosine” is a modified nucleobase. Auniversal base is a modified nucleobase that can pair with any one ofthe five unmodified nucleobases. As used herein, “nucleobase sequence”means the order of contiguous nucleobases in a target nucleic acid oroligonucleotide independent of any sugar or internucleoside linkagemodification.

As used herein, “nucleoside” means a compound, or a fragment of acompound, comprising a nucleobase and a sugar moiety. The nucleobase andsugar moiety are each, independently, unmodified or modified. As usedherein, “modified nucleoside” means a nucleoside comprising a modifiednucleobase and/or a modified sugar moiety. Modified nucleosides includeabasic nucleosides, which lack a nucleobase. “Linked nucleosides” arenucleosides that are connected in a contiguous sequence (i.e., noadditional nucleosides are presented between those that are linked).

As used herein, “oligomeric compound” means an oligonucleotide andoptionally one or more additional features, such as a conjugate group orterminal group. An oligomeric compound may be paired with a secondoligomeric compound that is complementary to the first oligomericcompound or may be unpaired. A “singled-stranded oligomeric compound” isan unpaired oligomeric compound. The term “oligomeric duplex” means aduplex formed by two oligomeric compounds having complementarynucleobase sequences. Each oligomeric compound of an oligomeric duplexmay be referred to as a “duplexed oligomeric compound.”

As used herein, “oligonucleotide” means a strand of linked nucleosidesconnected via internucleoside linkages, wherein each nucleoside andinternucleoside linkage may be modified or unmodified. Unless otherwiseindicated, oligonucleotides consist of 8-50 linked nucleosides. As usedherein, “modified oligonucleotide” means an oligonucleotide, wherein atleast one nucleoside or internucleoside linkage is modified. As usedherein, “unmodified oligonucleotide” means an oligonucleotide that doesnot comprise any nucleoside modifications or internucleosidemodifications.

As used herein, “pharmaceutically acceptable carrier or diluent” meansany substance suitable for use in administering to a subject. Certainsuch carriers enable pharmaceutical compositions to be formulated as,for example, tablets, pills, dragees, capsules, liquids, gels, syrups,slurries, suspension and lozenges for the oral ingestion by a subject.

In certain embodiments, a pharmaceutically acceptable carrier or diluentis sterile water, sterile saline, sterile buffer solution or sterileartificial cerebrospinal fluid.

As used herein, “pharmaceutically acceptable salts” meansphysiologically and pharmaceutically acceptable salts of compounds.Pharmaceutically acceptable salts retain the desired biological activityof the parent compound and do not impart undesired toxicological effectsthereto.

As used herein, “pharmaceutical composition” means a mixture ofsubstances suitable for administering to a subject. For example, apharmaceutical composition may comprise an oligomeric compound and asterile aqueous solution. In certain embodiments, a pharmaceuticalcomposition shows activity in free uptake assay in certain cell lines.

As used herein, “prodrug” means a therapeutic agent in a form outsidethe body that is converted to a different form within a subject or cellsthereof. Typically, conversion of a prodrug within the subject isfacilitated by the action of an enzymes (e.g., endogenous or viralenzyme) or chemicals present in cells or tissues and/or by physiologicconditions.

As used herein, “reducing the amount or activity” refers to a reductionor blockade of the transcriptional expression or activity relative tothe transcriptional expression or activity in an untreated or controlsample and does not necessarily indicate a total elimination oftranscriptional expression or activity.

As used herein, “RNA” means an RNA transcript and includes pre-mRNA andmature mRNA unless otherwise specified.

As used herein, “RNAi compound” means an antisense compound that acts,at least in part, through RISC or Ago2 to modulate a target nucleic acidand/or protein encoded by a target nucleic acid. RNAi compounds include,but are not limited to double-stranded siRNA, single-stranded RNA(ssRNA), and microRNA, including microRNA mimics. In certainembodiments, an RNAi compound modulates the amount, activity, and/orsplicing of a target nucleic acid. The term RNAi compound excludesantisense compounds that act through RNase H.

As used herein, “self-complementary” in reference to an oligonucleotidemeans an oligonucleotide that at least partially hybridizes to itself.

As used herein, “standard in vitro assay” means the assay described inExample 1 and reasonable variations thereof.

As used herein, “standard in vivo assay” means the assay described inExample 7 and reasonable variations thereof.

As used herein, “stereorandom chiral center” in the context of apopulation of molecules of identical molecular formula means a chiralcenter having a random stereochemical configuration. For example, in apopulation of molecules comprising a stereorandom chiral center, thenumber of molecules having the (S) configuration of the stereorandomchiral center may be but is not necessarily the same as the number ofmolecules having the (R) configuration of the stereorandom chiralcenter. The stereochemical configuration of a chiral center isconsidered random when it is the result of a synthetic method that isnot designed to control the stereochemical configuration. In certainembodiments, a stereorandom chiral center is a stereorandomphosphorothioate internucleoside linkage.

As used herein, “subject” means a human or non-human animal.

As used herein, “sugar moiety” means an unmodified sugar moiety or amodified sugar moiety. As used herein, “unmodified sugar moiety” means a2′-OH(H) β-D-ribosyl moiety, as found in RNA (an “unmodified RNA sugarmoiety”), or a 2′-H(H) β-D-deoxyribosyl sugar moiety, as found in DNA(an “unmodified DNA sugar moiety”). Unmodified sugar moieties have onehydrogen at each of the 1′, 3′, and 4′ positions, an oxygen at the 3′position, and two hydrogens at the 5′ position. As used herein,“modified sugar moiety” or “modified sugar” means a modified furanosylsugar moiety or a sugar surrogate.

As used herein, “sugar surrogate” means a modified sugar moiety havingother than a furanosyl moiety that can link a nucleobase to anothergroup, such as an internucleoside linkage, conjugate group, or terminalgroup in an oligonucleotide. Modified nucleosides comprising sugarsurrogates can be incorporated into one or more positions within anoligonucleotide and such oligonucleotides are capable of hybridizing tocomplementary oligomeric compounds or target nucleic acids.

As used herein, “symptom or hallmark” means any physical feature or testresult that indicates the existence or extent of a disease or disorder.In certain embodiments, a symptom is apparent to a subject or to amedical professional examining or testing said subject. In certainembodiments, a hallmark is apparent upon invasive diagnostic testing,including, but not limited to, post-mortem tests. In certainembodiments, a hallmark is apparent on a brain MRI scan.

As used herein, “target nucleic acid” and “target RNA” mean a nucleicacid that an antisense compound is designed to affect.

As used herein, “target region” means a portion of a target nucleic acidto which an oligomeric compound is designed to hybridize.

As used herein, “terminal group” means a chemical group or group ofatoms that is covalently linked to a terminus of an oligonucleotide.

As used herein, “therapeutically effective amount” means an amount of apharmaceutical agent that provides a therapeutic benefit to a subject.For example, a therapeutically effective amount improves a symptom orhallmark of a disease.

Certain Embodiments

The present disclosure provides the following non-limiting numberedembodiments:

-   -   Embodiment 1: An oligomeric compound comprising a modified        oligonucleotide consisting of 12 to 30 linked nucleosides        wherein the nucleobase sequence of the modified oligonucleotide        is at least 90% complementary to an equal length portion of a        GFAP nucleic acid, and wherein the modified oligonucleotide        comprises at least one modification selected from a modified        sugar moiety and a modified internucleoside linkage.    -   Embodiment 2: An oligomeric compound comprising a modified        oligonucleotide consisting of 12 to 30 linked nucleosides and        having a nucleobase sequence comprising at least 12, at least        13, at least 14, at least 15, at least 16, at least 17, at least        18, at least 19, or 20 contiguous nucleobases of any of SEQ ID        NOs: 20-2809 or 2813, wherein the modified oligonucleotide        comprises at least one modification selected from a modified        sugar moiety and a modified internucleoside linkage.    -   Embodiment 3: An oligomeric compound comprising a modified        oligonucleotide consisting of 12 to 30 linked nucleosides and        having a nucleobase sequence comprising at least 12, at least        13, at least 14, at least 15, at least 16, at least 17, or 18        contiguous nucleobases of any of SEQ ID NOs: 2816-2837,        2839-2846, 2850-2854, 2856, 2859, 2861-2863, 2866, 2873-2876,        2886-2888, 2891, wherein the modified oligonucleotide comprises        at least one modification selected from a modified sugar moiety        and a modified internucleoside linkage.    -   Embodiment 4: An oligomeric compound comprising a modified        oligonucleotide consisting of 12 to 30 linked nucleosides and        having a nucleobase sequence comprising at least 8, at least 9,        at least 10, at least 11, at least 12, at least 13, at least 14,        at least 15, at least 16, at least 17, at least 18, at least 19,        or at least 20 contiguous nucleobases complementary to:        -   an equal length portion of nucleobases 9324-9348 of SEQ ID            NO: 2;        -   an equal length portion of nucleobases 9459-9480 of SEQ ID            NO: 2;        -   an equal length portion of nucleobases 9530-9580 of SEQ ID            NO: 2;        -   an equal length portion of nucleobases 12006-12038 of SEQ ID            NO: 2; or        -   an equal length portion of nucleobases 13038-13058 of SEQ ID            NO: 2,        -   wherein the modified oligonucleotide comprises at least one            modification selected from a modified sugar moiety and a            modified internucleoside linkage.    -   Embodiment 5: An oligomeric compound comprising a modified        oligonucleotide consisting of 12 to 30 linked nucleosides and        having a nucleobase sequence comprising at least 8, at least 9,        at least 10, at least 11, at least 12, at least 13, at least 14,        at least 15, at least 16, at least 17, or at least 18 contiguous        nucleobases of a sequence selected from:        -   SEQ ID Nos: 21, 1177, 2321, 2398, 2808-2809, 2840-2842,            2853-2854;        -   SEQ ID Nos: 555, 2093, 2170, 2813;        -   SEQ ID Nos: 20, 88, 166, 1331, 1408, 1485, 1637, 1713, 1714,            1789, 1790, 1637, 1638, 1865, 1866, 1941, 2018, 2095, 2172,            2249, 2326, 2403, 2480, 2557, 2633, 2709, 2785, 2816-2818,            2859, 2861, 2886-2887;        -   SEQ ID Nos: 815, 893, 971, 1049, 1269, 1270, 1346, 1423,            1499, 1500, 1660, 1736, 2655, 2731; or        -   SEQ ID Nos: 825, 1973.    -   Embodiment 6: The oligomeric compound of any of embodiments 1-5,        wherein the modified oligonucleotide has a nucleobase sequence        that is at least 80%, at least 85%, at least 90%, at least 95%,        or 100% complementary to the nucleobase sequence of SEQ ID NO:        1, SEQ ID NO: 2 or SEQ ID NO: 3 when measured across the entire        nucleobase sequence of the modified oligonucleotide.    -   Embodiment 7: The oligomeric compound of any of embodiments 1-6,        wherein the modified oligonucleotide comprises at least one        modified sugar moiety.    -   Embodiment 8: The oligomeric compound of embodiment 7, wherein        the modified oligonucleotide comprises at least one bicyclic        sugar moiety.    -   Embodiment 9: The oligomeric compound of embodiment 8, wherein        the bicyclic sugar moiety has a 4′-2′ bridge, wherein the 4′-2′        bridge is selected from CH₂—O—; and —CH(CH₃)—O.    -   Embodiment 10: The oligomeric compound of any of embodiments        1-9, wherein the modified oligonucleotide comprises at least one        non-bicyclic modified sugar moiety.    -   Embodiment 11: The oligomeric compound of embodiment 10, wherein        the non-bicyclic modified sugar moiety is a 2′-MOE sugar moiety        or a 2′-OMe sugar moiety.    -   Embodiment 12: The oligomeric compound of any of embodiments        1-11, wherein the modified oligonucleotide comprises at least        one sugar surrogate.    -   Embodiment 13: The oligomeric compound of embodiment 12, wherein        the sugar surrogate is any of morpholino, modified morpholino,        PNA, THP, and F-HNA.    -   Embodiment 14: The oligomeric compound of any of embodiments 1-7        or 10-13, wherein the modified oligonucleotide does not comprise        a bicyclic sugar moiety.    -   Embodiment 15: The oligomeric compound of any of embodiments        1-14, wherein the modified oligonucleotide is a gapmer.    -   Embodiment 16: The oligomeric compound of any of embodiments        1-15, wherein the modified oligonucleotide comprises:    -   a 5′-region consisting of 1-6 linked nucleosides;    -   a central region consisting of 6-10 linked nucleosides; and a        3′-region consisting of 1-6 linked nucleosides; wherein each of        the 5′-region nucleosides and each of the 3′-region nucleosides        comprises a modified sugar moiety and at least one of the        central region nucleosides comprises a 2′-deoxyribosyl sugar        moiety.    -   Embodiment 17: The oligomeric compound of embodiment 16, wherein        the modified oligonucleotide comprises        -   a 5′-region consisting of 6 linked nucleosides;        -   a central region consisting of 10 linked nucleosides; and        -   a 3′-region consisting of 4 linked nucleosides; wherein        -   each of the 5′-region nucleosides and each of the 3′-region            nucleosides comprises a 2′-MOE modified sugar moiety, and            each of the central region nucleosides comprises a            2′-deoxyribosyl sugar moiety.    -   Embodiment 18: The oligomeric compound of embodiment 16, wherein        the modified oligonucleotide comprises:        -   a 5′-region consisting of 5 linked nucleosides;        -   a central region consisting of 10 linked nucleosides; and        -   a 3′-region consisting of 5 linked nucleosides; wherein        -   each of the 5′-region nucleosides and each of the 3′-region            nucleosides comprises a 2′-MOE modified sugar moiety, and            each of the central region nucleosides comprises a            2′-deoxyribosyl sugar moiety.    -   Embodiment 19: The oligomeric compound of embodiment 16, wherein        the modified oligonucleotide comprises:        -   a 5′-region consisting of 4 linked nucleosides;        -   a central region consisting of 10 linked nucleosides; and        -   a 3′-region consisting of 6 linked nucleosides; wherein        -   each of the 5′-region nucleosides and each of the 3′-region            nucleosides comprises a 2′-MOE modified sugar moiety, and            each of the central region nucleosides comprises a            2′-deoxyribosyl sugar moiety.    -   Embodiment 20: The oligomeric compound of embodiment 16, wherein        the modified oligonucleotide comprises        -   a 5′-region consisting of 5 linked nucleosides;        -   a central region consisting of 8 linked nucleosides; and        -   a 3′-region consisting of 5 linked nucleosides; wherein        -   each of the 5′-region nucleosides and each of the 3′-region            nucleosides comprises a 2′-MOE modified sugar moiety, and            each of the central region nucleosides comprises a            2′-deoxyribosyl sugar moiety.    -   Embodiment 21: The oligomeric compound of any of embodiments        1-20, wherein the modified oligonucleotide comprises at least        one modified internucleoside linkage.    -   Embodiment 22: The oligomeric compound of embodiment 21, wherein        each internucleoside linkage of the modified oligonucleotide is        a modified internucleoside linkage.    -   Embodiment 23: The oligomeric compound of embodiments 21 or 22,        wherein the modified internucleoside linkage is a        phosphorothioate internucleoside linkage.    -   Embodiment 24: The oligomeric compound of any one of embodiments        1-21, wherein the modified oligonucleotide comprises at least        one phosphodiester internucleoside linkage.    -   Embodiment 25: The oligomeric compound of any of embodiments 21,        23, or 24, wherein each internucleoside linkage is either a        phosphodiester internucleoside linkage or a phosphorothioate        internucleoside linkage.    -   Embodiment 26: The oligomeric compound of any one of embodiments        1-21 or 23-25, wherein the modified oligonucleotide has an        internucleoside linkage motif selected from among:        sooosssssssssssooss, sooooossssssssssoss, soooossssssssssooss,        sooosssssssssooss, or sooossssssssssoooss; wherein, s=a        phosphorothioate internucleoside linkage and o=a phosphodiester        internucleoside linkage.    -   Embodiment 27: The oligomeric compound of any of embodiments        1-26, wherein the modified oligonucleotide comprises at least        one modified nucleobase.    -   Embodiment 28: The oligomeric compound of embodiment 27, wherein        the modified nucleobase is a 5-methylcytosine.    -   Embodiment 29: The oligomeric compound of any of embodiments        1-28, wherein the modified oligonucleotide consists of 12-30,        12-22, 12-20, 14-18, 14-20, 15-17, 15-25, 16-20, 18-22 or 18-20        linked nucleosides.    -   Embodiment 30: The oligomeric compound of any of embodiments        1-29, wherein the modified oligonucleotide consists of 18 linked        nucleosides.    -   Embodiment 31: The oligomeric compound of any of embodiments        1-30, wherein the modified oligonucleotide consists of 20 linked        nucleosides.    -   Embodiment 32: The oligomeric compound of any of embodiments        1-31, consisting of the modified oligonucleotide.    -   Embodiment 33: An oligomeric duplex comprising an oligomeric        compound of any of embodiments 1-31.    -   Embodiment 34: An antisense compound comprising or consisting of        an oligomeric compound of any of embodiments 1-32 or an        oligomeric duplex of embodiment 33.    -   Embodiment 35: A pharmaceutical composition comprising an        oligomeric compound of any of embodiments 1-32 or an oligomeric        duplex of embodiment 33 and a pharmaceutically acceptable        carrier or diluent.    -   Embodiment 36: The pharmaceutical composition of embodiment 35,        wherein the pharmaceutically acceptable diluent is artificial        cerebral spinal fluid.    -   Embodiment 37: The pharmaceutical composition of embodiment 36,        wherein the pharmaceutical composition consists of the modified        oligonucleotide and phosphate buffered saline.    -   Embodiment 38: A method comprising administering to a subject a        pharmaceutical composition of any of embodiments 35-37.    -   Embodiment 39: A method of treating a disease associated with        GFAP comprising administering to an individual having or at risk        for developing a disease associated with GFAP a therapeutically        effective amount of a compound of any one of embodiments 1-34 or        a pharmaceutical composition according to any of embodiments        35-37; and thereby treating the disease associated with GFAP.    -   Embodiment 40: The method of embodiment 39, wherein the        GFAP-associated disease is Alexander Disease.    -   Embodiment 41: The method of any of embodiments 38-40, wherein        at least one symptom or hallmark of the GFAP-associated disease        is ameliorated.    -   Embodiment 42: The method of embodiment 41, wherein the symptom        or hallmark is motor delays, cognitive delays, paroxysmal        deterioration, seizures, vomiting, swallowing difficulties,        ataxic gait, palatal myoclonus, autonomic dysfunction, or the        presence of intra-astrocytic inclusions called Rosenthal fibers.    -   Embodiment 43: The method of any of embodiments 38-42, wherein        GFAP levels in the individual are reduced.    -   Embodiment 44: A modified oligonucleotide according to the        following chemical structure:

or a salt thereof.

-   -   Embodiment 45: A modified oligonucleotide according to the        following chemical structure:

-   -   Embodiment 46: A modified oligonucleotide according to the        following chemical structure:

or a salt thereof.

-   -   Embodiment 47: A modified oligonucleotide according to the        following chemical structure:

-   -   Embodiment 48: A modified oligonucleotide according to the        following chemical structure:

or a salt thereof.

-   -   Embodiment 49: A modified oligonucleotide according to the        following chemical structure:

-   -   Embodiment 50: A modified oligonucleotide according to the        following chemical structure:

or a salt thereof.

-   -   Embodiment 51: A modified oligonucleotide according to the        following chemical structure:

-   -   Embodiment 52: A modified oligonucleotide according to the        following chemical structure:

or a salt thereof.

-   -   Embodiment 53: A modified oligonucleotide according to the        following chemical structure:

-   -   Embodiment 54: A modified oligonucleotide according to the        following chemical structure:

or a salt thereof.

-   -   Embodiment 55: A modified oligonucleotide according to the        following chemical structure:

-   -   Embodiment 56: The modified oligonucleotide of any one of        embodiments 44, 46, 48, 50, 52, and 54, which is the sodium salt        or potassium salt of the chemical structure.    -   Embodiment 57: A pharmaceutical composition comprising the        modified oligonucleotide of any of embodiments 44-56 and a        pharmaceutically acceptable carrier or diluent.    -   Embodiment 58: The pharmaceutical composition of embodiment 57,        wherein the pharmaceutically acceptable diluent is artificial        cerebrospinal fluid.    -   Embodiment 59: The pharmaceutical composition of embodiment 58,        wherein the pharmaceutical composition consists of the modified        oligonucleotide and artificial cerebrospinal fluid.    -   Embodiment 60: A compound comprising a modified oligonucleotide        according to the following chemical notation:        ^(m)C_(es)A_(eo)G_(eo)T_(eo)A_(eo)T_(eo)T_(ds)A_(ds) ^(m)C_(as)        ^(m)C_(as)T_(ds) ^(m)C_(as)T_(ds)A_(ds)        ^(m)C_(as)T_(ds)A_(eo)G_(es)T_(es) ^(m)C_(e) (SEQ ID NO: 20),        wherein:    -   A=an adenine nucleobase,    -   ^(m)C=a 5-methylcytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-β-D-MOE sugar moiety,    -   d=a 2′-β-D-deoxyribosyl sugar moiety,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.    -   Embodiment 61: A compound comprising a modified oligonucleotide        according to the following chemical notation: ^(m)C_(es)A_(eo)        ^(m)C_(eo)A_(eo)T_(eo)T_(eo) ^(m)C_(ds)A_(ds)        ^(m)C_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)T_(ds)T_(ds)T_(eo)A_(es)A_(es)        ^(m)C_(e) (SEQ ID NO: 21), wherein:    -   A=an adenine nucleobase,    -   ^(m)C=a 5-methylcytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-β-D-MOE sugar moiety,    -   d=a 2′-β-D-deoxyribosyl sugar moiety,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.    -   Embodiment 62: A compound comprising a modified oligonucleotide        according to the following chemical notation: ^(m)C_(es)        ^(m)C_(eo)A_(eo)G_(eo)T_(eo)G_(ds)T_(ds) ^(m)C_(ds)T_(ds)T_(ds)        ^(m)C_(ds)A_(ds) ^(m)C_(ds)T_(ds)T_(ds)T_(eo)G_(eo)        ^(m)C_(es)T_(es) ^(m)C_(e) (SEQ ID NO: 825), wherein:    -   A=an adenine nucleobase,    -   ^(m)C=a 5-methylcytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-β-D-MOE sugar moiety,    -   d=a 2′-β-D-deoxyribosyl sugar moiety,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.    -   Embodiment 63: A compound comprising a modified oligonucleotide        according to the following chemical notation: G_(es)        ^(m)C_(eo)A_(eo)A_(eo) ^(m)C_(es)A_(ds)G_(ds)T_(ds)T_(ds)T_(ds)        ^(m)C_(ds) ^(m)C_(ds)A_(ds)T_(ds)A_(ds)A_(eo)        ^(m)C_(eo)A_(es)A_(es) ^(m)C_(e) (SEQ ID NO: 1499), wherein:    -   A=an adenine nucleobase,    -   ^(m)C=a 5-methylcytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-β-D-MOE sugar moiety,    -   d=a 2′-β-D-deoxyribosyl sugar moiety,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.    -   Embodiment 64: A compound comprising a modified oligonucleotide        according to the following chemical notation:        T_(es)G_(eo)G_(eo)T_(eo) ^(m)C_(eo)        ^(m)C_(ds)T_(ds)A_(ds)A_(ds)A_(ds)T_(ds)A_(ds)T_(ds)T_(ds)        ^(m)C_(ds)T_(eo)A_(eo)G_(es)T_(es) ^(m)C_(e) (SEQ ID NO: 2170),        wherein:    -   A=an adenine nucleobase,    -   ^(m)C=a 5-methylcytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-β-D-MOE sugar moiety,    -   d=a 2′-β-D-deoxyribosyl sugar moiety,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.    -   Embodiment 65: A compound comprising a modified oligonucleotide        according to the following chemical notation: T_(es)G_(eo)G_(eo)        ^(m)C_(eo)A_(es)G_(ds)T_(ds)A_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)        ^(m)C_(ds)T_(eo) ^(m)C_(eo)T_(es)A_(es) ^(m)C_(e) (SEQ ID NO:        2818), wherein:    -   A=an adenine nucleobase,    -   ^(m)C=a 5-methylcytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-β-D-MOE sugar moiety,    -   d=a 2′-β-D-deoxyribosyl sugar moiety,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.    -   Embodiment 66: The compound of any of embodiments 60-65,        comprising the modified oligonucleotide covalently linked to a        conjugate group.    -   Embodiment 67: A pharmaceutical composition comprising a        compound of any of embodiments 60-66, and a pharmaceutically        acceptable diluent or carrier.    -   Embodiment 68: The pharmaceutical composition of embodiment 67,        wherein the pharmaceutically acceptable diluent is artificial        cerebrospinal fluid.    -   Embodiment 69: The pharmaceutical composition of embodiment 68,        wherein the pharmaceutical composition consists of the compound        and artificial cerebrospinal fluid.    -   Embodiment 70: A chirally enriched population of modified        oligonucleotides of any of embodiments 60-65, wherein the        population is enriched for modified oligonucleotides comprising        at least one particular phosphorothioate internucleoside linkage        having a particular stereochemical configuration.    -   Embodiment 71: The chirally enriched population of embodiment        70, wherein the population is enriched for modified        oligonucleotides comprising at least one particular        phosphorothioate internucleoside linkage having the (Sp)        configuration.    -   Embodiment 72: The chirally enriched population of embodiment        70, wherein the population is enriched for modified        oligonucleotides comprising at least one particular        phosphorothioate internucleoside linkage having the (Rp)        configuration.    -   Embodiment 73: The chirally enriched population of embodiment        70, wherein the population is enriched for modified        oligonucleotides having a particular, independently selected        stereochemical configuration at each phosphorothioate        internucleoside linkage Embodiment 74: The chirally enriched        population of embodiment 70, wherein the population is enriched        for modified oligonucleotides having the (Sp) configuration at        each phosphorothioate internucleoside linkage.    -   Embodiment 75: The chirally enriched population of embodiment        70, wherein the population is enriched for modified        oligonucleotides having the (Rp) configuration at each        phosphorothioate internucleoside linkage.    -   Embodiment 76: The chirally enriched population of embodiment        70, wherein the population is enriched for modified        oligonucleotides having the (Rp) configuration at one particular        phosphorothioate internucleoside linkage and the (Sp)        configuration at each of the remaining phosphorothioate        internucleoside linkages.    -   Embodiment 77: The chirally enriched population of embodiment 70        or embodiment 73 wherein the population is enriched for modified        oligonucleotides having at least 3 contiguous phosphorothioate        internucleoside linkages in the Sp, Sp, and Rp configurations,        in the 5′ to 3′ direction.    -   Embodiment 78: A chirally enriched population of modified        oligonucleotides of any of embodiments 44-55, wherein all of the        phosphorothioate internucleoside linkages of the modified        oligonucleotide are stereorandom.    -   Embodiment 79: A method comprising administering to an        individual the pharmaceutical composition of any of embodiments        56-58 and 67-69.    -   Embodiment 80: A method of treating a disease associated with        GFAP, comprising administering to an individual having or at        risk of having a disease associated with GFAP a therapeutically        effective amount of the pharmaceutical composition of any one of        embodiments 56-58 and 67-69, thereby treating the disease        associated with GFAP.    -   Embodiment 81: A method of reducing GFAP protein in the CSF of        an individual having or at risk of having a disease associated        with GFAP a therapeutically effective amount of a pharmaceutical        composition of any one of embodiments 56-58 and 67-69, thereby        reducing GFAP protein in the CSF.    -   Embodiment 82: The method of embodiment 80 or embodiment 81,        wherein the disease is a neurodegenerative disease.    -   Embodiment 83: The method of any of embodiments 80-82, wherein        the disease is Alexander disease.    -   Embodiment 84: The method of any of embodiments 80-83, wherein        at least one symptom or hallmark of the disease is ameliorated.    -   Embodiment 85: The method of embodiment 84, wherein the symptom        or hallmark is any of motor delays, cognitive delays, paroxysmal        deterioration, seizures, vomiting, swallowing difficulties,        ataxic gait, palatal myoclonus, autonomic dysfunction, or the        presence of intra-astrocytic inclusions called Rosenthal fibers.    -   Embodiment 86: The method of any of embodiments 79-85, wherein        the pharmaceutical composition is administered to the central        nervous system or systemically.    -   Embodiment 87: The method of embodiment 86, wherein the        pharmaceutical composition is administered to the central        nervous system and systemically.    -   Embodiment 88: The method of any of embodiments 80-87, wherein        the pharmaceutical composition is administered any of        intrathecally, systemically, subcutaneously, or intramuscularly.

I. Certain Oligonucleotides

In certain embodiments, provided herein are oligomeric compoundscomprising oligonucleotides, which consist of linked nucleosides.Oligonucleotides may be unmodified oligonucleotides (RNA or DNA) or maybe modified oligonucleotides. Modified oligonucleotides comprise atleast one modification relative to unmodified RNA or DNA. That is,modified oligonucleotides comprise at least one modified nucleoside(comprising a modified sugar moiety and/or a modified nucleobase) and/orat least one modified internucleoside linkage.

A. Certain Modified Nucleosides

Modified nucleosides comprise a modified sugar moiety or a modifiednucleobase or both a modified sugar moiety and a modified nucleobase.

1. Certain Sugar Moieties

In certain embodiments, modified sugar moieties are non-bicyclicmodified sugar moieties. In certain embodiments, modified sugar moietiesare bicyclic or tricyclic sugar moieties. In certain embodiments,modified sugar moieties are sugar surrogates. Such sugar surrogates maycomprise one or more substitutions corresponding to those of other typesof modified sugar moieties.

In certain embodiments, modified sugar moieties are non-bicyclicmodified sugar moieties comprising a furanosyl ring with one or moresubstituent groups none of which bridges two atoms of the furanosyl ringto form a bicyclic structure. Such non bridging substituents may be atany position of the furanosyl, including but not limited to substituentsat the 2′, 4′, and/or 5′ positions. In certain embodiments one or morenon-bridging substituent of non-bicyclic modified sugar moieties isbranched. Examples of 2′-substituent groups suitable for non-bicyclicmodified sugar moieties include but are not limited to: 2′-F, 2

OCH₃ (“OMe” or “O-methyl”), and 2′-O(CH₂)₂OCH₃ (“MOE” or“O-methoxyethyl”). In certain embodiments, 2′-substituent groups areselected from among: halo, allyl, amino, azido, SH, CN, OCN, CF₃, OCF₃,O—C₁-C₁₀ alkoxy, O—C₁-C₁₀ substituted alkoxy, 0-C₁-C₁₀ alkyl, O—C₁-C₁₀substituted alkyl, S-alkyl, N(R_(m))-alkyl, O-alkenyl, S-alkenyl,N(R_(m))-alkenyl, O-alkynyl, S-alkynyl, N(R_(m))-alkynyl,O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl,O(CH₂)₂SCH₃, O(CH₂)₂ON(R_(m))(R_(n)) or OCH₂C(═O)—N(R_(m))(R_(n)), whereeach R_(m) and R_(n) is, independently, H, an amino protecting group, orsubstituted or unsubstituted C₁-C₁₀ alkyl, and the 2′-substituent groupsdescribed in Cook et al., U.S. Pat. No. 6,531,584; Cook et al., U.S.Pat. No. 5,859,221; and Cook et al., U.S. Pat. No. 6,005,087. Certainembodiments of these 2

substituent groups can be further substituted with one or moresubstituent groups independently selected from among: hydroxyl, amino,alkoxy, carboxy, benzyl, phenyl, nitro (NO₂), thiol, thioalkoxy,thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl. Examples of4′-substituent groups suitable for non-bicyclic modified sugar moietiesinclude but are not limited to alkoxy (e.g., methoxy), alkyl, and thosedescribed in Manoharan et al., WO 2015/106128. Examples of5′-substituent groups suitable for non-bicyclic modified sugar moietiesinclude but are not limited to: 5′-methyl (R or S), 5

vinyl, and 5′-methoxy. In certain embodiments, non-bicyclic modifiedsugar moieties comprise more than one non-bridging sugar substituent,for example, 2

5

methyl sugar moieties and the modified sugar moieties and modifiednucleosides described in Migawa et al., WO 2008/101157 and Rajeev etal., US2013/0203836.

In certain embodiments, a 2′-substituted non-bicyclic modifiednucleoside comprises a sugar moiety comprising a non-bridging2′-substituent group selected from: F, NH₂, N₃, OCF₃, OCH₃, O(CH₂)₃NH₂,CH₂CH═CH₂, OCH₂CH═CH₂, OCH₂CH₂OCH₃, O(CH₂)₂SCH₃,O(CH₂)₂ON(R_(m))(R_(n)), O(CH₂)₂O(CH₂)₂N(CH₃)₂, and N-substitutedacetamide (OCH₂C(═O)—N(R_(m))(R_(n))), where each R_(m) and R_(n) is,independently, H, an amino protecting group, or substituted orunsubstituted C₁-C₁₀ alkyl.

In certain embodiments, a 2′-substituted non-bicyclic modifiednucleoside comprises a sugar moiety comprising a non-bridging2′-substituent group selected from: F, OCF₃, OCH₃, OCH₂CH₂OCH₃,O(CH₂)₂SCH₃, O(CH₂)₂ON(CH₃)₂, O(CH₂)₂O(CH₂)₂N(CH₃)₂, andOCH₂C(═O)—N(H)CH₃ (“NMA”).

In certain embodiments, a 2′-substituted non-bicyclic modifiednucleoside comprises a sugar moiety comprising a non-bridging2′-substituent group selected from: F, OCH₃, and OCH₂CH₂OCH₃.

In certain embodiments, modified furanosyl sugar moieties andnucleosides incorporating such modified furanosyl sugar moieties arefurther defined by isomeric configuration. For example, a2′-deoxyfuranosyl sugar moiety may be in seven isomeric configurationsother than the naturally occurring β-D-deoxyribosyl configuration. Suchmodified sugar moieties are described in, e.g., WO 2019/157531,incorporated by reference herein. A 2′-modified sugar moiety has anadditional stereocenter at the 2′-position relative to a2′-deoxyfuranosyl sugar moiety; therefore, such sugar moieties have atotal of sixteen possible isomeric configurations. 2′-modified sugarmoieties described herein are in the β-D-ribosyl isomeric configurationunless otherwise specified.

Certain modified sugar moieties comprise a substituent that bridges twoatoms of the furanosyl ring to form a second ring, resulting in abicyclic sugar moiety. In certain such embodiments, the bicyclic sugarmoiety comprises a bridge between the 4′ and the 2′ furanose ring atoms.Examples of such 4′ to 2′ bridging sugar substituents include but arenot limited to: 4

CH₂-2

4

(CH₂)₂-2

4

(CH₂)₃-2

4

CH₂—O-2′ (“LNA”), 4′-CH₂—S-2

4

CH₂)₂—O-2′ (“ENA”), 4′-CH(CH₃)—O-2′ (referred to as “constrained ethyl”or “cEt”), 4′-CH₂—O—CH₂-2′, 4′-CH₂—N(R)-2′, 4′-CH(CH₂OCH₃)—O-2□(“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth etal., U.S. Pat. No. 7,399,845, Bhat et al., U.S. Pat. No. 7,569,686,Swayze et al., U.S. Pat. No. 7,741,457, and Swayze et al., U.S. Pat. No.8,022,193), 4

C(CH₃)(CH₃)—O-2□ and analogs thereof (see, e.g., Seth et al., U.S. Pat.No. 8,278,283), 4

CH₂—N(OCH₃)-2□ and analogs thereof (see, e.g., Prakash et al., U.S. Pat.No. 8,278,425), 4

CH₂—O—N(CH₃)-2□ (see, e.g., Allerson et al., U.S. Pat. No. 7,696,345 andAllerson et al., U.S. Pat. No. 8,124,745), 4

CH₂—C(H)(CH₃)-2□ (see, e.g., Zhou, et al., J. Org. Chem., 2009, 74,118-134), 4

CH₂—C(═CH₂)-2□ and analogs thereof (see e.g., Seth et al., U.S. Pat. No.8,278,426), 4′-C(R_(a)R_(b))—N(R)—O-2′, 4′-C(R_(a)R_(b))—O—N(R)-2′, 4

CH₂—O—N(R)-2

and 4

CH₂—N(R)—O-2

wherein each R, R_(a), and R_(b) is, independently, H, a protectinggroup, or C₁-C₁₂ alkyl (see, e.g. Imanishi et al., U.S. Pat. No.7,427,672).

In certain embodiments, such 4′ to 2′ bridges independently comprisefrom 1 to 4 linked groups independently selected from:—[C(R_(a))(R_(b))]_(n)—, —[C(R_(a))(R_(b))]_(n)—O—, —C(R_(a))═C(R_(b))—,—C(R_(a))═N—, —C(═NR_(a))—, —C(═O)—, —C(═S)—, —O—, —Si(R_(a))₂—,—S(═O)_(x)—, and —N(R_(a))—;

wherein:

x is 0, 1, or 2;

n is 1, 2, 3, or 4;

each R_(a) and R_(b) is, independently, H, a protecting group, hydroxyl,C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substitutedC₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl,substituted C₅-C₂₀ aryl, heterocycle radical, substituted heterocycleradical, heteroaryl, substituted heteroaryl, C₅-C₇ alicyclic radical,substituted C₅-C₇ alicyclic radical, halogen, OJ₁, NJ₁J₂, SJ₁, N₃,COOJ₁, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)₂-J₁), orsulfoxyl (S(═O)-J₁); and

each J₁ and J₂ is, independently, H, C₁-C₁₂ alkyl, substituted C₁-C₁₂alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl,substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, acyl(C(═O)—H), substituted acyl, a heterocycle radical, a substitutedheterocycle radical, C₁-C₁₂ aminoalkyl, substituted C₁-C₁₂ aminoalkyl,or a protecting group.

Additional bicyclic sugar moieties are known in the art, see, forexample: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443,Albaek et al., J. Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem.Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54,3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222;Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al.,J. Am. Chem. Soc., 2007, 129, 8362-8379; Wengel et al., U.S. Pat. No.7,053,207; Imanishi et al., U.S. Pat. No. 6,268,490; Imanishi et al.U.S. Pat. No. 6,770,748; Imanishi et al., U.S. RE44,779; Wengel et al.,U.S. Pat. No. 6,794,499; Wengel et al., U.S. Pat. No. 6,670,461; Wengelet al., U.S. Pat. No. 7,034,133; Wengel et al., U.S. Pat. No. 8,080,644;Wengel et al., U.S. Pat. No. 8,034,909; Wengel et al., U.S. Pat. No.8,153,365; Wengel et al., U.S. Pat. No. 7,572,582; Ramasamy et al., U.S.Pat. No. 6,525,191; Torsten et al., WO 2004/106356; Wengel et al., WO1999/014226; Seth et al., WO 2007/134181; Seth et al., U.S. Pat. No.7,547,684; Seth et al., U.S. Pat. No. 7,666,854; Seth et al., U.S. Pat.No. 8,088,746; Seth et al., U.S. Pat. No. 7,750,131; Seth et al., U.S.Pat. No. 8,030,467; Seth et al., U.S. Pat. No. 8,268,980; Seth et al.,U.S. Pat. No. 8,546,556; Seth et al., U.S. Pat. No. 8,530,640; Migawa etal., U.S. Pat. No. 9,012,421; Seth et al., U.S. Pat. No. 8,501,805; andU.S. Patent Publication Nos. Allerson et al., US2008/0039618 and Migawaet al., US2015/0191727.

In certain embodiments, bicyclic sugar moieties and nucleosidesincorporating such bicyclic sugar moieties are further defined byisomeric configuration. For example, an LNA nucleoside (describedherein) may be in the α-L configuration or in the β-D configuration.

α-L-methyleneoxy (4′-CH₂—O-2′) or α-L-LNA bicyclic nucleosides have beenincorporated into oligonucleotides that showed antisense activity(Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). Herein,general descriptions of bicyclic nucleosides include both isomericconfigurations. When the positions of specific bicyclic nucleosides(e.g., LNA or cEt) are identified in exemplified embodiments herein,they are in the β-D configuration, unless otherwise specified.

In certain embodiments, modified sugar moieties comprise one or morenon-bridging sugar substituent and one or more bridging sugarsubstituent (e.g., 5′-substituted and 4′-2′ bridged sugars).

In certain embodiments, modified sugar moieties are sugar surrogates. Incertain such embodiments, the oxygen atom of the sugar moiety isreplaced, e.g., with a sulfur, carbon or nitrogen atom. In certain suchembodiments, such modified sugar moieties also comprise bridging and/ornon-bridging substituents as described herein. For example, certainsugar surrogates comprise a 4′-sulfur atom and a substitution at the 2

position (see, e.g., Bhat et al., U.S. Pat. No. 7,875,733 and Bhat etal., U.S. Pat. No. 7,939,677) and/or the 5′ position.

In certain embodiments, sugar surrogates comprise rings having otherthan 5 atoms. For example, in certain embodiments, a sugar surrogatecomprises a six-membered tetrahydropyran (“THP”). Such tetrahydropyransmay be further modified or substituted. Nucleosides comprising suchmodified tetrahydropyrans include but are not limited to hexitol nucleicacid (“HNA”), anitol nucleic acid (“ANA”), manitol nucleic acid (“MNA”)(see, e.g., Leumann, C J. Bioorg. &Med. Chem. 2002, 10, 841-854), fluoroHNA:

(“F-HNA”, see e.g. Swayze et al., U.S. Pat. No. 8,088,904; Swayze etal., U.S. Pat. No. 8,440,803; Swayze et al., U.S. Pat. No. 8,796,437;and Swayze et al., U.S. Pat. No. 9,005,906; F-HNA can also be referredto as a F-THP or 3

fluoro tetrahydropyran), and nucleosides comprising additional modifiedTHP compounds having the formula:

wherein, independently, for each of the modified THP nucleosides:

Bx is a nucleobase moiety;

T₃ and T₄ are each, independently, an internucleoside linking grouplinking the modified THP nucleoside to the remainder of anoligonucleotide or one of T₃ and T₄ is an internucleoside linking grouplinking the modified THP nucleoside to the remainder of anoligonucleotide and the other of T₃ and T₄ is H, a hydroxyl protectinggroup, a linked conjugate group, or a 5□ or 3

terminal group;

q₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each, independently, H, C₁-C₆ alkyl,substituted C₁-C₆ alkyl, C₂-C₆ alkenyl, substituted C₂-C₆ alkenyl, C₂-C₆alkynyl, or substituted C₂-C₆ alkynyl; and

each of R₁ and R₂ is independently selected from among: hydrogen,halogen, substituted or unsubstituted alkoxy, NJ₁J₂, SJ₁, N₃, OC(═X)J₁,OC(═X)NJ₁J₂, NJ₃C(═X)NJ₁J₂, and CN, wherein X is O, S or NJ₁, and eachJ₁, J₂, and J₃ is, independently, H or C₁-C₆ alkyl.

In certain embodiments, modified THP nucleosides are provided whereinq₁, q₂, q₃, q₄, q₅, q₆ and gp are each H. In certain embodiments, atleast one of q₁, q₂, q₃, q₄, q₅, q₆ and gp is other than H. In certainembodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆ and gp is methyl. Incertain embodiments, modified THP nucleosides are provided wherein oneof R₁ and R₂ is F. In certain embodiments, R₁ is F and R₂ is H, incertain embodiments, R₁ is methoxy and R₂ is H, and in certainembodiments, R₁ is methoxyethoxy and R₂ is H.

In certain embodiments, sugar surrogates comprise rings having more than5 atoms and more than one heteroatom. For example, nucleosidescomprising morpholino sugar moieties and their use in oligonucleotideshave been reported (see, e.g., Braasch et al., Biochemistry, 2002, 41,4503-4510 and Summerton et al., U.S. Pat. No. 5,698,685; Summerton etal., U.S. Pat. No. 5,166,315; Summerton et al., U.S. Pat. No. 5,185,444;and Summerton et al., U.S. Pat. No. 5,034,506). As used here, the term“morpholino” means a sugar surrogate having the following structure:

In certain embodiments, morpholinos may be modified, for example byadding or altering various substituent groups from the above morpholinostructure. Such sugar surrogates are referred to herein as “modifiedmorpholinos.”

In certain embodiments, sugar surrogates comprise acyclic moieties.Examples of nucleosides and oligonucleotides comprising such acyclicsugar surrogates include but are not limited to: peptide nucleic acid(“PNA”), acyclic butyl nucleic acid (see, e.g., Kumar et al., Org.Biomol. Chem., 2013, 11, 5853-5865), and nucleosides andoligonucleotides described in Manoharan et al., WO2011/133876.

Many other bicyclic and tricyclic sugar and sugar surrogate ring systemsare known in the art that can be used in modified nucleosides.

2. Certain Modified Nucleobases

In certain embodiments, modified oligonucleotides comprise one or morenucleosides comprising an unmodified nucleobase. In certain embodiments,modified oligonucleotides comprise one or more nucleoside comprising amodified nucleobase. In certain embodiments, modified oligonucleotidescomprise one or more nucleoside that does not comprise a nucleobase,referred to as an abasic nucleoside.

In certain embodiments, modified nucleobases are selected from:5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynylsubstituted pyrimidines, alkyl substituted purines, and N-2, N-6 and 0-6substituted purines. In certain embodiments, modified nucleobases areselected from: 2-aminopropyladenine, 5-hydroxymethyl cytosine, xanthine,hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine,2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine,5-propynyl (—C≡C—CH₃) uracil, 5-propynylcytosine, 6-azouracil,6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil),4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-azaand other 8-substituted purines, 5-halo, particularly 5-bromo,5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine,7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine,7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine,2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases,hydrophobic bases, promiscuous bases, size-expanded bases, andfluorinated bases. Further modified nucleobases include tricyclicpyrimidines, such as 1,3-diazaphenoxazine-2-one,1,3-diazaphenothiazine-2-one and9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp). Modifiednucleobases may also include those in which the purine or pyrimidinebase is replaced with other heterocycles, for example 7-deaza-adenine,7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobasesinclude those disclosed in Merigan et al., U.S. Pat. No. 3,687,808,those disclosed in The Concise Encyclopedia Of Polymer Science AndEngineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859;Englisch et al., Angewandte Chemie, International Edition, 1991, 30,613; Sanghvi, Y. S., Chapter 15, Antisense Research and Applications,Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and thosedisclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S. T.,Ed., CRC Press, 2008, 163-166 and 442-443.

Publications that teach the preparation of certain of the above notedmodified nucleobases as well as other modified nucleobases includewithout limitation, Manoharan et al., US2003/0158403; Manoharan et al.,US2003/0175906; Dinh et al., U.S. Pat. No. 4,845,205; Spielvogel et al.,U.S. Pat. No. 5,130,302; Rogers et al., U.S. Pat. No. 5,134,066;Bischofberger et al., U.S. Pat. No. 5,175,273; Urdea et al., U.S. Pat.No. 5,367,066; Benner et al., U.S. Pat. No. 5,432,272; Matteucci et al.,U.S. Pat. No. 5,434,257; Gmeiner et al., U.S. Pat. No. 5,457,187; Cooket al., U.S. Pat. No. 5,459,255; Froehler et al., U.S. Pat. No.5,484,908; Matteucci et al., U.S. Pat. No. 5,502,177; Hawkins et al.,U.S. Pat. No. 5,525,711; Haralambidis et al., U.S. Pat. No. 5,552,540;Cook et al., U.S. Pat. No. 5,587,469; Froehler et al., U.S. Pat. No.5,594,121; Switzer et al., U.S. Pat. No. 5,596,091; Cook et al., U.S.Pat. No. 5,614,617; Froehler et al., U.S. Pat. No. 5,645,985; Cook etal., U.S. Pat. No. 5,681,941; Cook et al., U.S. Pat. No. 5,811,534; Cooket al., U.S. Pat. No. 5,750,692; Cook et al., U.S. Pat. No. 5,948,903;Cook et al., U.S. Pat. No. 5,587,470; Cook et al., U.S. Pat. No.5,457,191; Matteucci et al., U.S. Pat. No. 5,763,588; Froehler et al.,U.S. Pat. No. 5,830,653; Cook et al., U.S. Pat. No. 5,808,027; Cook etal., 6,166,199; and Matteucci et al., U.S. Pat. No. 6,005,096.

3. Certain Modified Internucleoside Linkages

In certain embodiments, nucleosides of modified oligonucleotides may belinked together using any internucleoside linkage. The two main classesof internucleoside linking groups are defined by the presence or absenceof a phosphorus atom. Representative phosphorus-containinginternucleoside linkages include but are not limited to phosphodiesters,which contain a phosphodiester bond (“P(O₂)═O”) (also referred to asunmodified or naturally occurring linkages), phosphotriesters,methylphosphonates, phosphoramidates, phosphorothioates (“P(O₂)═S”), andphosphorodithioates (“HS-P═S”). Representative non-phosphorus containinginternucleoside linking groups include but are not limited tomethylenemethylimino (—CH₂—N(CH₃)—O—CH₂—), thiodiester, thionocarbamate(—O—C(═O)(NH)—S—); siloxane (—O—SiH₂—O—); and N,N

dimethylhydrazine (—CH₂—N(CH₃)—N(CH₃)—). Modified internucleosidelinkages, compared to naturally occurring phosphodiester internucleosidelinkages, can be used to alter, typically increase, nuclease resistanceof the oligonucleotide. In certain embodiments, internucleoside linkageshaving a chiral atom can be prepared as a racemic mixture, or asseparate enantiomers. Methods of preparation of phosphorous-containingand non-phosphorous-containing internucleoside linkages are well knownto those skilled in the art.

Representative internucleoside linkages having a chiral center includebut are not limited to alkylphosphonates and phosphorothioates. Modifiedoligonucleotides comprising internucleoside linkages having a chiralcenter can be prepared as populations of modified oligonucleotidescomprising stereorandom internucleoside linkages, or as populations ofmodified oligonucleotides comprising phosphorothioate internucleosidelinkages in particular stereochemical configurations. In certainembodiments, populations of modified oligonucleotides comprisephosphorothioate internucleoside linkages wherein all of thephosphorothioate internucleoside linkages are stereorandom. Suchmodified oligonucleotides can be generated using synthetic methods thatresult in random selection of the stereochemical configuration of eachphosphorothioate internucleoside linkage. Nonetheless, as is wellunderstood by those of skill in the art, each individualphosphorothioate of each individual oligonucleotide molecule has adefined stereoconfiguration. In certain embodiments, populations ofmodified oligonucleotides are enriched for modified oligonucleotidescomprising one or more particular phosphorothioate internucleosidelinkage in a particular, independently selected stereochemicalconfiguration. In certain embodiments, the particular configuration ofthe particular phosphorothioate internucleoside linkage is present in atleast 65% of the molecules in the population. In certain embodiments,the particular configuration of the particular phosphorothioateinternucleoside linkage is present in at least 70% of the molecules inthe population. In certain embodiments, the particular configuration ofthe particular phosphorothioate internucleoside linkage is present in atleast 80% of the molecules in the population. In certain embodiments,the particular configuration of the particular phosphorothioateinternucleoside linkage is present in at least 90% of the molecules inthe population. In certain embodiments, the particular configuration ofthe particular phosphorothioate internucleoside linkage is present in atleast 99% of the molecules in the population. Such chirally enrichedpopulations of modified oligonucleotides can be generated usingsynthetic methods known in the art, e.g., methods described in Oka etal., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res. 42, 13456 (2014),and WO 2017/015555. In certain embodiments, a population of modifiedoligonucleotides is enriched for modified oligonucleotides having atleast one indicated phosphorothioate in the (Sp) configuration. Incertain embodiments, a population of modified oligonucleotides isenriched for modified oligonucleotides having at least onephosphorothioate in the (Rp) configuration. In certain embodiments,modified oligonucleotides comprising (Rp) and/or (Sp) phosphorothioatescomprise one or more of the following formulas, respectively, wherein“B” indicates a nucleobase:

Unless otherwise indicated, chiral internucleoside linkages of modifiedoligonucleotides described herein can be stereorandom or in a particularstereochemical configuration.

Neutral internucleoside linkages include, without limitation,phosphotriesters, methylphosphonates, MMI (3

CH₂—N(CH₃)—O-5

, amide-3 (3

CH₂—C(═O)—N(H)-5

, amide-4 (3

CH₂—N(H)—C(═O)-5

, formacetal (3

O—CH₂—O-5

, methoxypropyl (MOP), and thioformacetal (3

S—CH₂—O-5

. Further neutral internucleoside linkages include nonionic linkagescomprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide,sulfide, sulfonate ester and amides (see for example: CarbohydrateModifications in Antisense Research; Y. S. Sanghvi and P. D. Cook, Eds.,ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutralinternucleoside linkages include nonionic linkages comprising mixed N,O, S and CH₂ component parts.

B. Certain Motifs

In certain embodiments, modified oligonucleotides comprise one or moremodified nucleosides comprising a modified sugar moiety. In certainembodiments, modified oligonucleotides comprise one or more modifiednucleosides comprising a modified nucleobase. In certain embodiments,modified oligonucleotides comprise one or more modified internucleosidelinkage. In such embodiments, the modified, unmodified, and differentlymodified sugar moieties, nucleobases, and/or internucleoside linkages ofa modified oligonucleotide define a pattern or motif. In certainembodiments, the patterns of sugar moieties, nucleobases, andinternucleoside linkages are each independent of one another. Thus, amodified oligonucleotide may be described by its sugar motif, nucleobasemotif and/or internucleoside linkage motif (as used herein, nucleobasemotif describes the modifications to the nucleobases independent of thesequence of nucleobases).

1. Certain Sugar Motifs

In certain embodiments, oligonucleotides comprise one or more type ofmodified sugar and/or unmodified sugar moiety arranged along theoligonucleotide or portion thereof in a defined pattern or sugar motif.In certain instances, such sugar motifs include but are not limited toany of the sugar modifications discussed herein.

In certain embodiments, modified oligonucleotides have a gapmer motif,which is defined by two external regions or “wings” and a central orinternal region or “gap.” The three regions of a gapmer motif (the5′-wing, the gap, and the 3′-wing) form a contiguous sequence ofnucleosides wherein at least some of the sugar moieties of thenucleosides of each of the wings differ from at least some of the sugarmoieties of the nucleosides of the gap. Specifically, at least the sugarmoieties of the nucleosides of each wing that are closest to the gap(the 3′-most nucleoside of the 5′-wing and the 5′-most nucleoside of the3′-wing) differ from the sugar moiety of the neighboring gapnucleosides, thus defining the boundary between the wings and the gap(i.e., the wing/gap junction). In certain embodiments, the sugarmoieties within the gap are the same as one another. In certainembodiments, the gap includes one or more nucleoside having a sugarmoiety that differs from the sugar moiety of one or more othernucleosides of the gap. In certain embodiments, the sugar motifs of thetwo wings are the same as one another (symmetric gapmer). In certainembodiments, the sugar motif of the 5

wing differs from the sugar motif of the 3

wing (asymmetric gapmer).

In certain embodiments, the wings of a gapmer comprise 1-6 nucleosides.In certain embodiments, each nucleoside of each wing of a gapmercomprises a modified sugar moiety. In certain embodiments, at least onenucleoside of each wing of a gapmer comprises a modified sugar moiety.In certain embodiments, at least two nucleosides of each wing of agapmer comprises a modified sugar moiety. In certain embodiments, atleast three nucleosides of each wing of a gapmer comprises a modifiedsugar moiety. In certain embodiments, at least four nucleosides of eachwing of a gapmer comprises a modified sugar moiety. In certainembodiments, at least five nucleosides of each wing of a gapmercomprises a modified sugar moiety.

In certain embodiments, the gap of a gapmer comprises 7-12 nucleosides.In certain embodiments, each nucleoside of the gap of a gapmer comprisesa 2-deoxyribosyl sugar moiety. In certain embodiments, each nucleosideof the gap of a gapmer comprises a 2′-β-D-deoxyribosyl sugar moiety. Incertain embodiments, at least one nucleoside of the gap of a gapmercomprises a modified sugar moiety. In certain embodiments, at least onenucleoside of the gap of a gapmer comprises a 2′-OMe sugar moiety.

In certain embodiments, the gapmer is a deoxy gapmer. In certainembodiments, the nucleosides on the gap side of each wing/gap junctioncomprise 2′-deoxyribosyl sugar moieties and the nucleosides on the wingsides of each wing/gap junction comprise modified sugar moieties. Incertain embodiments, each nucleoside of the gap comprises a2-deoxyribosyl sugar moiety. In certain embodiments, each nucleoside ofeach wing of a gapmer comprises a modified sugar moiety. In certainembodiments, one nucleoside of the gap comprises a modified sugar moietyand each remaining nucleoside of the gap comprises a 2-deoxyribosylsugar moiety.

In certain embodiments, modified oligonucleotides comprise or consist ofa portion having a fully modified sugar motif. In such embodiments, eachnucleoside of the fully modified portion of the modified oligonucleotidecomprises a modified sugar moiety. In certain embodiments, eachnucleoside of the entire modified oligonucleotide comprises a modifiedsugar moiety. In certain embodiments, modified oligonucleotides compriseor consist of a portion having a fully modified sugar motif, whereineach nucleoside within the fully modified portion comprises the samemodified sugar moiety, referred to herein as a uniformly modified sugarmotif. In certain embodiments, a fully modified oligonucleotide is auniformly modified oligonucleotide. In certain embodiments, eachnucleoside of a uniformly modified oligonucleotide comprises the same2′-modification.

Herein, the lengths (number of nucleosides) of the three regions of agapmer may be provided using the notation [# of nucleosides in the5′-wing]−[# of nucleosides in the gap]−[# of nucleosides in the3′-wing]. Thus, a 5-10-5 gapmer consists of 5 linked nucleosides in eachwing and 10 linked nucleosides in the gap. Where such nomenclature isfollowed by a specific modification, that modification is themodification in each sugar moiety of each wing and the gap nucleosidescomprises a 2′-β-D-deoxyribosyl sugar moiety. Thus, a 5-10-5 MOE gapmerconsists of 5 linked 2′-MOE nucleosides in the 5′-wing, 10 linked2′-β-D-deoxynucleosides in the gap, and 5 linked 2′-MOE nucleosides inthe 3′-wing. A 3-10-3 cEt gapmer consists of 3 linked cEt nucleosides inthe 5′-wing, 10 linked 2′-β-D-deoxynucleosides in the gap, and 3 linkedcEt nucleosides in the 3′-wing. A 5-8-5 gapmer consists of 5 linkednucleosides comprising a modified sugar moiety in the 5′-wing, 8 linked2′-deoxynucleosides in the gap, and 5 linked nucleosides comprising amodified sugar moiety in the 3′-wing. A mixed wing gapmer has at leasttwo different modified sugars in the 5′ and/or 3′ wing. A 5-8-5 or 5-8-4mixed wing gapmer has at least two different modified sugar moieties inthe 5′- and/or the 3′-wing.

In certain embodiments, modified oligonucleotides are 5-10-5 MOEgapmers. In certain embodiments, modified oligonucleotides are 4-10-6MOE gapmers. In certain embodiments, modified oligonucleotides are6-10-4 MOE gapmers. In certain embodiments, modified oligonucleotidesare 5-8-5 MOE gapmers. In certain embodiments, modified oligonucleotidesare X-Y-Z MOE gapmers, wherein X and Z are independently selected from1, 2, 3, 4, 5, or 6 and Y is 7, 8, 9, 10, or 11.

In certain embodiments, modified oligonucleotides have the followingsugar motif (5′ to 3′): meeemddddddddddmmmmm, wherein ‘d’ represents a2′-deoxyribosyl sugar moiety, ‘e’ represents a 2′-MOE sugar moiety, and‘m’ represents a 2′-OMe sugar moiety.

2. Certain Nucleobase Motifs

In certain embodiments, oligonucleotides comprise modified and/orunmodified nucleobases arranged along the oligonucleotide or portionthereof in a defined pattern or motif. In certain embodiments, eachnucleobase is modified. In certain embodiments, none of the nucleobasesare modified. In certain embodiments, each purine or each pyrimidine ismodified. In certain embodiments, each adenine is modified. In certainembodiments, each guanine is modified. In certain embodiments, eachthymine is modified. In certain embodiments, each uracil is modified. Incertain embodiments, each cytosine is modified. In certain embodiments,some or all of the cytosine nucleobases in a modified oligonucleotideare 5-methylcytosines. In certain embodiments, all of the cytosinenucleobases are 5-methylcytosines and all of the other nucleobases ofthe modified oligonucleotide are unmodified nucleobases.

In certain embodiments, modified oligonucleotides comprise a block ofmodified nucleobases. In certain such embodiments, the block is at the3′-end of the oligonucleotide. In certain embodiments the block iswithin 3 nucleosides of the 3′-end of the oligonucleotide. In certainembodiments, the block is at the 5′-end of the oligonucleotide. Incertain embodiments the block is within 3 nucleosides of the 5′-end ofthe oligonucleotide.

In certain embodiments, oligonucleotides having a gapmer motif comprisea nucleoside comprising a modified nucleobase. In certain suchembodiments, one nucleoside comprising a modified nucleobase is in thecentral gap of an oligonucleotide having a gapmer motif. In certain suchembodiments, the sugar moiety of the nucleoside is a 2′-deoxyribosylsugar moiety. In certain embodiments, the modified nucleobase isselected from: a 2-thiopyrimidine and a 5-propynepyrimidine.

3. Certain Internucleoside Linkage Motifs

In certain embodiments, oligonucleotides comprise modified and/orunmodified internucleoside linkages arranged along the oligonucleotideor region thereof in a defined pattern or motif. In certain embodiments,each internucleoside linking group is a phosphodiester internucleosidelinkage (P═O). In certain embodiments, each internucleoside linkinggroup of a modified oligonucleotide is a phosphorothioateinternucleoside linkage (P═S). In certain embodiments, eachinternucleoside linkage of a modified oligonucleotide is independentlyselected from a phosphorothioate internucleoside linkage andphosphodiester internucleoside linkage. In certain embodiments, eachphosphorothioate internucleoside linkage is independently selected froma stereorandom phosphorothioate, a (Sp) phosphorothioate, and a (Rp)phosphorothioate. In certain embodiments, the sugar motif of a modifiedoligonucleotide is a gapmer and the internucleoside linkages within thegap are all modified. In certain such embodiments, some or all of theinternucleoside linkages in the wings are unmodified phosphodiesterinternucleoside linkages. In certain embodiments, the terminalinternucleoside linkages are modified. In certain embodiments, the sugarmotif of a modified oligonucleotide is a gapmer, and the internucleosidelinkage motif comprises at least one phosphodiester internucleosidelinkage in at least one wing, wherein the at least one phosphodiesterinternucleoside linkage is not a terminal internucleoside linkage, andthe remaining internucleoside linkages are phosphorothioateinternucleoside linkages. In certain such embodiments, all of thephosphorothioate internucleoside linkages are stereorandom. In certainembodiments, all of the phosphorothioate internucleoside linkages in thewings are (Sp) phosphorothioates, and the gap comprises at least one Sp,Sp, Rp motif. In certain embodiments, populations of modifiedoligonucleotides are enriched for modified oligonucleotides comprisingsuch internucleoside linkage motifs.

In certain embodiments, modified oligonucleotides have aninternucleoside linkage motif of sooosssssssssssooss, wherein each “s”represents a phosphorothioate internucleoside linkage and each “o”represents a phosphodiester internucleoside linkage. In certainembodiments, modified oligonucleotides have an internucleoside linkagemotif of (5′ to 3′): sooooossssssssssoss, wherein each “s” represents aphosphorothioate internucleoside linkage and each “o” represents aphosphodiester internucleoside linkage. In certain embodiments, modifiedoligonucleotides have an internucleoside linkage motif of (5′ to 3′):soooossssssssssooss, wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage. In certain embodiments, modifiedoligonucleotides have an internucleoside linkage motif of (5′ to3′):sooosssssssssooss, wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage. In certain embodiments, modifiedoligonucleotides have an internucleoside linkage motif of (5′ to 3′):sooossssssssssoooss, wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage. In certain embodiments, modifiedoligonucleotides have an internucleoside linkage motif of (5′ to 3′):sooosssssssssssssss, wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage.

C. Certain Lengths

It is possible to increase or decrease the length of an oligonucleotidewithout eliminating activity. For example, in Woolf et al. Proc. Natl.Acad. Sci. USA 89:7305-7309, 1992) a series of oligonucleotides 13-25nucleobases in length were tested for their ability to induce cleavageof a target nucleic acid in an oocyte injection model. Oligonucleotides25 nucleobases in length with 8 or 11 mismatch bases near the ends ofthe oligonucleotides were able to direct specific cleavage of the targetnucleic acid, albeit to a lesser extent than the oligonucleotides thatcontained no mismatches. Similarly, target specific cleavage wasachieved using 13 nucleobase oligonucleotides, including those with 1 or3 mismatches.

In certain embodiments, oligonucleotides (including modifiedoligonucleotides) can have any of a variety of ranges of lengths. Incertain embodiments, oligonucleotides consist of X to Y linkednucleosides, where X represents the fewest number of nucleosides in therange and Y represents the largest number nucleosides in the range. Incertain such embodiments, X and Y are each independently selected from8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, and 50; provided that X<Y. For example, incertain embodiments, oligonucleotides consist of 12 to 13, 12 to 14, 12to 15, 12 to 16, 12 to 17, 12 to 18, 12 to 19, 12 to 20, 12 to 21, 12 to22, 12 to 23, 12 to 24, 12 to 25, 12 to 26, 12 to 27, 12 to 28, 12 to29, 12 to 30, 13 to 14, 13 to 15, 13 to 16, 13 to 17, 13 to 18, 13 to19, 13 to 20, 13 to 21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to26, 13 to 27, 13 to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29, 16 to30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to24, 17 to 25, 17 to 26, 17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25, 18 to26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 29, 19 to 28, 19 to29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to26, 20 to 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29, 21 to 30, 22 to23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to30, 23 to 24, 23 to 25, 23 to 26, 23 to 27, 23 to 28, 23 to 29, 23 to30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to30 linked nucleosides.

D. Certain Modified Oligonucleotides

In certain embodiments, the above modifications (sugar, nucleobase,internucleoside linkage) are incorporated into a modifiedoligonucleotide. In certain embodiments, modified oligonucleotides arecharacterized by their modification motifs and overall lengths. Incertain embodiments, such parameters are each independent of oneanother. Thus, unless otherwise indicated, each internucleoside linkageof an oligonucleotide having a gapmer sugar motif may be modified orunmodified and may or may not follow the gapmer modification pattern ofthe sugar modifications. For example, the internucleoside linkageswithin the wing regions of a sugar gapmer may be the same or differentfrom one another and may be the same or different from theinternucleoside linkages of the gap region of the sugar motif. Likewise,such sugar gapmer oligonucleotides may comprise one or more modifiednucleobase independent of the gapmer pattern of the sugar modifications.Unless otherwise indicated, all modifications are independent ofnucleobase sequence.

E. Certain Populations of Modified Oligonucleotides

Populations of modified oligonucleotides in which all of the modifiedoligonucleotides of the population have the same molecular formula canbe stereorandom populations or chirally enriched populations. All of thechiral centers of all of the modified oligonucleotides are stereorandomin a stereorandom population. In a chirally enriched population, atleast one particular chiral center is not stereorandom in the modifiedoligonucleotides of the population. In certain embodiments, the modifiedoligonucleotides of a chirally enriched population are enriched for β-Dribosyl sugar moieties, and all of the phosphorothioate internucleosidelinkages are stereorandom. In certain embodiments, the modifiedoligonucleotides of a chirally enriched population are enriched for bothβ-D ribosyl sugar moieties and at least one, particular phosphorothioateinternucleoside linkage in a particular stereochemical configuration.

F. Nucleobase Sequence

In certain embodiments, oligonucleotides (unmodified or modifiedoligonucleotides) are further described by their nucleobase sequence. Incertain embodiments oligonucleotides have a nucleobase sequence that iscomplementary to a second oligonucleotide or an identified referencenucleic acid, such as a target nucleic acid. In certain suchembodiments, a portion of an oligonucleotide has a nucleobase sequencethat is complementary to a second oligonucleotide or an identifiedreference nucleic acid, such as a target nucleic acid. In certainembodiments, the nucleobase sequence of a portion or entire length of anoligonucleotide is at least 50%, at least 60%, at least 70%, at least80%, at least 85%, at least 90%, at least 95%, or 100% complementary tothe second oligonucleotide or nucleic acid, such as a target nucleicacid.

II. Certain Oligomeric Compounds

In certain embodiments, provided herein are oligomeric compounds, whichconsist of an oligonucleotide (modified or unmodified) and optionallyone or more conjugate groups and/or terminal groups. Conjugate groupsconsist of one or more conjugate moiety and a conjugate linker whichlinks the conjugate moiety to the oligonucleotide. Conjugate groups maybe attached to either or both ends of an oligonucleotide and/or at anyinternal position. In certain embodiments, conjugate groups are attachedto the 2 position of a nucleoside of a modified oligonucleotide. Incertain embodiments, conjugate groups that are attached to either orboth ends of an oligonucleotide are terminal groups. In certain suchembodiments, conjugate groups or terminal groups are attached at the 3′and/or 5′-end of oligonucleotides. In certain such embodiments,conjugate groups (or terminal groups) are attached at the 3′-end ofoligonucleotides. In certain embodiments, conjugate groups are attachednear the 3′-end of oligonucleotides. In certain embodiments, conjugategroups (or terminal groups) are attached at the 5′-end ofoligonucleotides. In certain embodiments, conjugate groups are attachednear the 5′-end of oligonucleotides.

Examples of terminal groups include but are not limited to conjugategroups, capping groups, phosphate moieties, protecting groups, abasicnucleosides, modified or unmodified nucleosides, and two or morenucleosides that are independently modified or unmodified.

A. Certain Conjugate Groups

In certain embodiments, oligonucleotides are covalently attached to oneor more conjugate groups. In certain embodiments, conjugate groupsmodify one or more properties of the attached oligonucleotide, includingbut not limited to pharmacodynamics, pharmacokinetics, stability,binding, absorption, tissue distribution, cellular distribution,cellular uptake, charge and clearance. In certain embodiments, conjugategroups impart a new property on the attached oligonucleotide, e.g.,fluorophores or reporter groups that enable detection of theoligonucleotide. Certain conjugate groups and conjugate moieties havebeen described previously, for example: cholesterol moiety (Letsinger etal., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid(Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), athioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad.Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett.,1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. AcidsRes., 1992, 20, 533-538), an aliphatic chain, e.g., do-decan-diol orundecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10, 1111-1118;Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al.,Biochimie, 1993, 75, 49-54), a phospholipid, e.g.,di-hexadecyl-rac-glycerol or triethyl-ammonium1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al.,Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res.,1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain(Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), oradamantane acetic acid a palmityl moiety (Mishra et al., Biochim.Biophys. Acta, 1995, 1264, 229-237), an octadecylamine orhexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol.Exp. Ther., 1996, 277, 923-937), a tocopherol group (Nishina et al.,Molecular Therapy Nucleic Acids, 2015, 4, e220; and Nishina et al.,Molecular Therapy, 2008, 16, 734-740), or a GalNAc cluster (e.g.,WO2014/179620).

In certain embodiments, conjugate groups may be selected from any of aC22 alkyl, C20 alkyl, C16 alkyl, C10 alkyl, C21 alkyl, C19 alkyl, C18alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, C11 alkyl, C9 alkyl,C8 alkyl, C7 alkyl, C6 alkyl, C5 alkyl, C22 alkenyl, C20 alkenyl, C16alkenyl, C10 alkenyl, C21 alkenyl, C19 alkenyl, C18 alkenyl, C15alkenyl, C14 alkenyl, C13 alkenyl, C12 alkenyl, C11 alkenyl, C9 alkenyl,C8 alkenyl, C7 alkenyl, C6 alkenyl, or C5 alkenyl.

In certain embodiments, conjugate groups may be selected from any of C22alkyl, C20 alkyl, C16 alkyl, C10 alkyl, C21 alkyl, C19 alkyl, C18 alkyl,C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, C11 alkyl, C9 alkyl, C8alkyl, C7 alkyl, C6 alkyl, and C5 alkyl, where the alkyl chain has oneor more unsaturated bonds.

1. Conjugate Moieties

Conjugate moieties include, without limitation, intercalators, reportermolecules, polyamines, polyamides, peptides, carbohydrates, vitaminmoieties, polyethylene glycols, thioethers, polyethers, cholesterols,thiocholesterols, cholic acid moieties, folate, lipids, lipophilicgroups, phospholipids, biotin, phenazine, phenanthridine, anthraquinone,adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores,and dyes.

In certain embodiments, a conjugate moiety comprises an active drugsubstance, for example, aspirin, warfarin, phenylbutazone, ibuprofen,suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen,dansylsarcosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid,folinic acid, a benzothiadiazide, chlorothiazide, a diazepine,indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, anantidiabetic, an antibacterial or an antibiotic.

2. Conjugate Linkers

Conjugate moieties are attached to oligonucleotides through conjugatelinkers. In certain oligomeric compounds, the conjugate linker is asingle chemical bond (i.e., the conjugate moiety is attached directly toan oligonucleotide through a single bond). In certain embodiments, theconjugate linker comprises a chain structure, such as a hydrocarbylchain, or an oligomer of repeating units such as ethylene glycol,nucleosides, or amino acid units.

In certain embodiments, a conjugate linker comprises one or more groupsselected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol,ether, thioether, and hydroxylamino. In certain such embodiments, theconjugate linker comprises groups selected from alkyl, amino, oxo, amideand ether groups. In certain embodiments, the conjugate linker comprisesgroups selected from alkyl and amide groups. In certain embodiments, theconjugate linker comprises groups selected from alkyl and ether groups.In certain embodiments, the conjugate linker comprises at least onephosphorus moiety. In certain embodiments, the conjugate linkercomprises at least one phosphate group. In certain embodiments, theconjugate linker includes at least one neutral linking group.

In certain embodiments, conjugate linkers, including the conjugatelinkers described above, are bifunctional linking moieties, e.g., thoseknown in the art to be useful for attaching conjugate groups to parentcompounds, such as the oligonucleotides provided herein. In general, abifunctional linking moiety comprises at least two functional groups.One of the functional groups is selected to bind to a particular site ona parent compound and the other is selected to bind to a conjugategroup. Examples of functional groups used in a bifunctional linkingmoiety include but are not limited to electrophiles for reacting withnucleophilic groups and nucleophiles for reacting with electrophilicgroups. In certain embodiments, bifunctional linking moieties compriseone or more groups selected from amino, hydroxyl, carboxylic acid,thiol, alkyl, alkenyl, and alkynyl.

Examples of conjugate linkers include but are not limited topyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include butare not limited to substituted or unsubstituted C₁-C₁₀ alkyl,substituted or unsubstituted C₂-C₁₀ alkenyl or substituted orunsubstituted C₂-C₁₀ alkynyl, wherein a nonlimiting list of preferredsubstituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl,phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl andalkynyl.

In certain embodiments, conjugate linkers comprise 1-10linker-nucleosides. In certain embodiments, conjugate linkers comprise2-5 linker-nucleosides. In certain embodiments, conjugate linkerscomprise exactly 3 linker-nucleosides. In certain embodiments, conjugatelinkers comprise the TCA motif. In certain embodiments, suchlinker-nucleosides are modified nucleosides. In certain embodiments suchlinker-nucleosides comprise a modified sugar moiety. In certainembodiments, linker-nucleosides are unmodified. In certain embodiments,linker-nucleosides comprise an optionally protected heterocyclic baseselected from a purine, substituted purine, pyrimidine or substitutedpyrimidine. In certain embodiments, a cleavable moiety is a nucleosideselected from uracil, thymine, cytosine, 4-N-benzoylcytosine,5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine,6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is typicallydesirable for linker-nucleosides to be cleaved from the oligomericcompound after it reaches a target tissue. Accordingly,linker-nucleosides are typically linked to one another and to theremainder of the oligomeric compound through cleavable bonds. In certainembodiments, such cleavable bonds are phosphodiester bonds.

Herein, linker-nucleosides are not considered to be part of theoligonucleotide. Accordingly, in embodiments in which an oligomericcompound comprises an oligonucleotide consisting of a specified numberor range of linked nucleosides and/or a specified percentcomplementarity to a reference nucleic acid and the oligomeric compoundalso comprises a conjugate group comprising a conjugate linkercomprising linker-nucleosides, those linker-nucleosides are not countedtoward the length of the oligonucleotide and are not used in determiningthe percent complementarity of the oligonucleotide for the referencenucleic acid. For example, an oligomeric compound may comprise (1) amodified oligonucleotide consisting of 8-30 nucleosides and (2) aconjugate group comprising 1-10 linker-nucleosides that are contiguouswith the nucleosides of the modified oligonucleotide. The total numberof contiguous linked nucleosides in such an oligomeric compound is morethan 30. Alternatively, an oligomeric compound may comprise a modifiedoligonucleotide consisting of 8-30 nucleosides and no conjugate group.The total number of contiguous linked nucleosides in such an oligomericcompound is no more than 30. Unless otherwise indicated conjugatelinkers comprise no more than 10 linker-nucleosides. In certainembodiments, conjugate linkers comprise no more than 5linker-nucleosides. In certain embodiments, conjugate linkers compriseno more than 3 linker-nucleosides. In certain embodiments, conjugatelinkers comprise no more than 2 linker-nucleosides. In certainembodiments, conjugate linkers comprise no more than 1linker-nucleoside.

In certain embodiments, it is desirable for a conjugate group to becleaved from the oligonucleotide. For example, in certain circumstancesoligomeric compounds comprising a particular conjugate moiety are bettertaken up by a particular cell type, but once the oligomeric compound hasbeen taken up, it is desirable that the conjugate group be cleaved torelease the unconjugated or parent oligonucleotide. Thus, certainconjugate linkers may comprise one or more cleavable moieties. Incertain embodiments, a cleavable moiety is a cleavable bond. In certainembodiments, a cleavable moiety is a group of atoms comprising at leastone cleavable bond. In certain embodiments, a cleavable moiety comprisesa group of atoms having one, two, three, four, or more than fourcleavable bonds. In certain embodiments, a cleavable moiety isselectively cleaved inside a cell or subcellular compartment, such as alysosome. In certain embodiments, a cleavable moiety is selectivelycleaved by endogenous enzymes, such as nucleases.

In certain embodiments, a cleavable bond is selected from among: anamide, an ester, an ether, one or both esters of a phosphodiester, aphosphate ester, a carbamate, or a disulfide. In certain embodiments, acleavable bond is one or both of the esters of a phosphodiester. Incertain embodiments, a cleavable moiety comprises a phosphate orphosphodiester. In certain embodiments, the cleavable moiety is aphosphate or phosphodiester internucleoside linkage between anoligonucleotide and a conjugate moiety or conjugate group.

In certain embodiments, a cleavable moiety comprises or consists of oneor more linker-nucleosides. In certain such embodiments, the one or morelinker-nucleosides are linked to one another and/or to the remainder ofthe oligomeric compound through cleavable bonds. In certain embodiments,such cleavable bonds are unmodified phosphodiester bonds. In certainembodiments, a cleavable moiety is 2

deoxynucleoside that is attached to either the 3□ or 5

terminal nucleoside of an oligonucleotide by a phosphodiesterinternucleoside linkage and covalently attached to the remainder of theconjugate linker or conjugate moiety by a phosphate or phosphorothioateinternucleoside linkage.

In certain such embodiments, the cleavable moiety is 2

deoxyadenosine.

3. Cell-Targeting Moieties

In certain embodiments, a conjugate group comprises a cell-targetingmoiety. In certain embodiments, a conjugate group has the generalformula:

wherein n is from 1 to about 3, m is 0 when n is 1, m is 1 when n is 2or greater, j is 1 or 0, and k is 1 or 0.

In certain embodiments, n is 1, j is 1 and k is 0. In certainembodiments, n is 1, j is 0 and k is 1. In certain embodiments, n is 1,j is 1 and k is 1. In certain embodiments, n is 2, j is 1 and k is 0. Incertain embodiments, n is 2, j is 0 and k is 1. In certain embodiments,n is 2, j is 1 and k is 1. In certain embodiments, n is 3, j is 1 and kis 0. In certain embodiments, n is 3, j is 0 and k is 1. In certainembodiments, n is 3, j is 1 and k is 1.

In certain embodiments, conjugate groups comprise cell-targetingmoieties that have at least one tethered ligand. In certain embodiments,cell-targeting moieties comprise two tethered ligands covalentlyattached to a branching group. In certain embodiments, cell-targetingmoieties comprise three tethered ligands covalently attached to abranching group.

B. Certain Terminal Groups

In certain embodiments, oligomeric compounds comprise one or moreterminal groups. In certain such embodiments, oligomeric compoundscomprise a stabilized 5′-phosphate. Stabilized 5′-phosphates include,but are not limited to 5′-phosphonates, including, but not limited to5′-vinylphosphonates. In certain embodiments, terminal groups compriseone or more abasic nucleosides and/or inverted nucleosides. In certainembodiments, terminal groups comprise one or more 2′-linked nucleosides.In certain such embodiments, the 2′-linked nucleoside is an abasicnucleoside.

III. Oligomeric Duplexes

In certain embodiments, oligomeric compounds described herein comprisean oligonucleotide, having a nucleobase sequence complementary to thatof a target nucleic acid. In certain embodiments, an oligomeric compoundis paired with a second oligomeric compound to form an oligomericduplex. Such oligomeric duplexes comprise a first oligomeric compoundhaving a portion complementary to a target nucleic acid and a secondoligomeric compound having a portion complementary to the firstoligomeric compound. In certain embodiments, the first oligomericcompound of an oligomeric duplex comprises or consists of (1) a modifiedor unmodified oligonucleotide and optionally a conjugate group and (2) asecond modified or unmodified oligonucleotide and optionally a conjugategroup. Either or both oligomeric compounds of an oligomeric duplex maycomprise a conjugate group. The oligonucleotides of each oligomericcompound of an oligomeric duplex may include non-complementaryoverhanging nucleosides.

IV. Antisense Activity

In certain embodiments, oligomeric compounds and oligomeric duplexes arecapable of hybridizing to a target nucleic acid, resulting in at leastone antisense activity; such oligomeric compounds and oligomericduplexes are antisense compounds. In certain embodiments, antisensecompounds have antisense activity when they reduce the amount oractivity of a target nucleic acid by 25% or more in the standard cellassay. In certain embodiments, antisense compounds selectively affectone or more target nucleic acid. Such antisense compounds comprise anucleobase sequence that hybridizes to one or more target nucleic acid,resulting in one or more desired antisense activity and does nothybridize to one or more non-target nucleic acid or does not hybridizeto one or more non-target nucleic acid in such a way that results insignificant undesired antisense activity.

In certain antisense activities, hybridization of an antisense compoundto a target nucleic acid results in recruitment of a protein thatcleaves the target nucleic acid. For example, certain antisensecompounds result in RNase H mediated cleavage of the target nucleicacid. RNase H is a cellular endonuclease that cleaves the RNA strand ofan RNA:DNA duplex. The DNA in such an RNA:DNA duplex need not beunmodified DNA. In certain embodiments, described herein are antisensecompounds that are sufficiently “DNA-like” to elicit RNase H activity.In certain embodiments, one or more non-DNA-like nucleoside in the gapof a gapmer is tolerated.

In certain antisense activities, an antisense compound or a portion ofan antisense compound is loaded into an RNA-induced silencing complex(RISC), ultimately resulting in cleavage of the target nucleic acid. Forexample, certain antisense compounds result in cleavage of the targetnucleic acid by Argonaute. Antisense compounds that are loaded into RISCare RNAi compounds. RNAi compounds may be double-stranded (siRNA) orsingle-stranded (ssRNA).

In certain embodiments, hybridization of an antisense compound to atarget nucleic acid does not result in recruitment of a protein thatcleaves that target nucleic acid. In certain embodiments, hybridizationof the antisense compound to the target nucleic acid results inalteration of splicing of the target nucleic acid. In certainembodiments, hybridization of an antisense compound to a target nucleicacid results in inhibition of a binding interaction between the targetnucleic acid and a protein or other nucleic acid. In certainembodiments, hybridization of an antisense compound to a target nucleicacid results in alteration of translation of the target nucleic acid.

Antisense activities may be observed directly or indirectly. In certainembodiments, observation or detection of an antisense activity involvesobservation or detection of a change in an amount of a target nucleicacid or protein encoded by such target nucleic acid, a change in theratio of splice variants of a nucleic acid or protein and/or aphenotypic change in a cell or subject.

V. Certain Target Nucleic Acids

In certain embodiments, oligomeric compounds comprise or consist of anoligonucleotide comprising a portion that is complementary to a targetnucleic acid. In certain embodiments, the target nucleic acid is anendogenous RNA molecule. In certain embodiments, the target nucleic acidencodes a protein. In certain such embodiments, the target nucleic acidis selected from: a mature mRNA and a pre-mRNA, including intronic,exonic and untranslated regions. In certain embodiments, the targetnucleic acid is a mature mRNA. In certain embodiments, the targetnucleic acid is a pre-mRNA. In certain embodiments, the target region isentirely within an intron. In certain embodiments, the target regionspans an intron/exon junction. In certain embodiments, the target regionis at least 50% within an intron.

A. Complementarity/Mismatches to the Target Nucleic Acid

It is possible to introduce mismatch bases without eliminating activity.For example, Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March2001) demonstrated the ability of an oligonucleotide having 100%complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xLmRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and invivo. Furthermore, this oligonucleotide demonstrated potent anti-tumoractivity in vivo. Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988)tested a series of tandem 14 nucleobase oligonucleotides, and a 28- and42-nucleobase oligonucleotides comprised of the sequence of two or threeof the tandem oligonucleotides, respectively, for their ability toarrest translation of human DHFR in a rabbit reticulocyte assay. Each ofthe three 14 nucleobase oligonucleotides alone was able to inhibittranslation, albeit at a more modest level than the 28 or 42 nucleobaseoligonucleotides.

In certain embodiments, oligonucleotides are complementary to the targetnucleic acid over the entire length of the oligonucleotide. In certainembodiments, oligonucleotides are 99%, 95%, 90%, 85%, or 80%complementary to the target nucleic acid. In certain embodiments,oligonucleotides are at least 80% complementary to the target nucleicacid over the entire length of the oligonucleotide and comprise aportion that is 100% or fully complementary to a target nucleic acid. Incertain embodiments, the portion of full complementarity is 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24nucleobases in length.

In certain embodiments, oligonucleotides comprise one or more mismatchednucleobases relative to the target nucleic acid. In certain embodiments,antisense activity against the target is reduced by such mismatch, butactivity against a non-target is reduced by a greater amount. Thus, incertain embodiments selectivity of the oligonucleotide is improved. Incertain embodiments, the mismatch is specifically positioned within anoligonucleotide having a gapmer motif. In certain embodiments, themismatch is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 from the5′-end of the gap region. In certain embodiments, the mismatch is atposition 1, 2, 3, 4, 5, or 6 from the 5′-end of the 5′ wing region orthe 3′ wing region.

B. GFAP

In certain embodiments, oligomeric compounds comprise or consist of anoligonucleotide that is complementary to a target nucleic acid, whereinthe target nucleic acid is a GFAP nucleic acid. In certain embodiments,GFAP nucleic acid has the sequence set forth in SEQ ID NO: 1 (GENBANKAccession No. NM_002055.4), SEQ ID NO: 2 (GENBANK Accession No.NC_000017.11 truncated from nucleotides 44903001 to 44919000), or SEQ IDNO: 3 (GENBANK Accession No. NM_001131019.2).

In certain embodiments, contacting a cell with an oligomeric compoundcomplementary to any of SEQ ID NO: 1-3 reduces the amount of GFAP RNAand in certain embodiments reduces the amount of GFAP protein. Incertain embodiments, the oligomeric compound consists of a modifiedoligonucleotide. In certain embodiments, contacting a cell with anoligomeric compound complementary to any of SEQ ID NO: 1-3 reduces theamount of GFAP RNA in a cell, and in certain embodiments reduces theamount of GFAP protein in a cell. In certain embodiments, the cell is invitro. In certain embodiments, the cell is in a subject. In certainembodiments, the oligomeric compound consists of a modifiedoligonucleotide. In certain embodiments, contacting a cell in a subjectwith an oligomeric compound complementary to any of SEQ ID NO: 1-3ameliorates one or more symptom or hallmark of a leukodystrophy. Incertain embodiments, the leukodystrophy is AxD. In certain embodiments,the symptom or hallmark is selected from motor delays, cognitive delays,paroxysmal deterioration, seizures, vomiting, swallowing difficulties,ataxic gait, palatal myoclonus, autonomic dysfunction, and presence ofintra-astrocytic inclusions called Rosenthal fibers.

In certain embodiments, an oligomeric compound complementary to any ofSEQ ID NO: 1-3 is capable of reducing the detectable amount of GFAP RNAin vitro by at least 10%, at least 20%, at least 30%, at least 40%, atleast 50%, at least 60%, at least 70%, at least 80%, or at least 90%when administered according to the standard cell assay. In certainembodiments, an oligomeric compound complementary to SEQ ID NO: 1, SEQID NO: 2915, or SEQ ID NO: 2916 is capable of decreasing the amount ofGFAP in vitro by at least 10%, at least 20%, at least 30%, at least 40%,at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%when administered according to the standard in vitro assay. In certainembodiments, an oligomeric compound complementary to SEQ ID NO: 1, SEQID NO: 2, or SEQ ID NO: 3 is capable of reducing the detectable amountof GFAP RNA in the CSF of a subject by at least 10%, at least 20%, atleast 30%, at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, or at least 90%. In certain embodiments, an oligomericcompound complementary to SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 iscapable of decreasing the detectable amount of GFAP in the CSF of asubject by at least 10%, at least 20%, at least 30%, at least 40%, atleast 50%, at least 60%, at least 70%, at least 80%, or at least 90%.

C. Certain Target Nucleic Acids in Certain Tissues

In certain embodiments, oligomeric compounds comprise or consist of anoligonucleotide comprising a portion that is complementary to a targetnucleic acid, wherein the target nucleic acid is expressed in apharmacologically relevant tissue. In certain embodiments, thepharmacologically relevant tissues are the cells and tissues thatcomprise the central nervous system (CNS). Such tissues include thebrain and spinal cord. In certain embodiments, the pharmacologicallyrelevant tissues include white matter tracts across the brain and spinalcord, such tissues include the corpus callosum, cortex, cerebellum,hippocampus, brain stem, striatum, and spinal cord. In certainembodiments, the pharmacologically relevant tissues include the cortex,cerebellum, hippocampus, brain stem, and spinal cord. In certainembodiments, the pharmacologically relevant cells are oligodendrocytesand oligodendrocyte progenitor cells. In certain embodiments, thepharmacologically relevant cells are Schwann cells or Schwann cellprogenitors.

VI. Certain Pharmaceutical Compositions

In certain embodiments, described herein are pharmaceutical compositionscomprising one or more oligomeric compounds. In certain embodiments, theone or more oligomeric compounds each consists of a modifiedoligonucleotide. In certain embodiments, the pharmaceutical compositioncomprises a pharmaceutically acceptable diluent or carrier. In certainembodiments, a pharmaceutical composition comprises or consists of asterile saline solution and one or more oligomeric compound. In certainembodiments, the sterile saline is pharmaceutical grade saline. Incertain embodiments, a pharmaceutical composition comprises or consistsof one or more oligomeric compound and sterile water. In certainembodiments, the sterile water is pharmaceutical grade water. In certainembodiments, a pharmaceutical composition comprises or consists of oneor more oligomeric compound and phosphate-buffered saline (PBS). Incertain embodiments, the sterile PBS is pharmaceutical grade PBS. Incertain embodiments, a pharmaceutical composition comprises or consistsof one or more oligomeric compound and artificial cerebrospinal fluid(“artificial CSF” or “aCSF”). In certain embodiments, the artificialcerebrospinal fluid is pharmaceutical grade.

In certain embodiments, a pharmaceutical composition comprises amodified oligonucleotide and artificial cerebrospinal fluid. In certainembodiments, a pharmaceutical composition consists of a modifiedoligonucleotide and artificial cerebrospinal fluid. In certainembodiments, a pharmaceutical composition consists essentially of amodified oligonucleotide and artificial cerebrospinal fluid. In certainembodiments, the artificial cerebrospinal fluid is pharmaceutical grade.

In certain embodiments, pharmaceutical compositions comprise one or moreoligomeric compound and one or more excipients. In certain embodiments,excipients are selected from water, salt solutions, alcohol,polyethylene glycols, gelatin, lactose, amylase, magnesium stearate,talc, silicic acid, viscous paraffin, hydroxymethylcellulose andpolyvinylpyrrolidone.

In certain embodiments, oligomeric compounds may be admixed withpharmaceutically acceptable active and/or inert substances for thepreparation of pharmaceutical compositions or formulations. Compositionsand methods for the formulation of pharmaceutical compositions depend ona number of criteria, including, but not limited to, route ofadministration, extent of disease, or dose to be administered.

In certain embodiments, pharmaceutical compositions comprising anoligomeric compound encompass any pharmaceutically acceptable salts ofthe oligomeric compound, esters of the oligomeric compound, or salts ofsuch esters. In certain embodiments, pharmaceutical compositionscomprising oligomeric compounds comprising one or more oligonucleotide,upon administration to a subject, including a human, are capable ofproviding (directly or indirectly) the biologically active metabolite orresidue thereof. Accordingly, for example, the disclosure is also drawnto pharmaceutically acceptable salts of oligomeric compounds, prodrugs,pharmaceutically acceptable salts of such prodrugs, and otherbioequivalents. Suitable pharmaceutically acceptable salts include, butare not limited to, sodium and potassium salts. In certain embodiments,prodrugs comprise one or more conjugate group attached to anoligonucleotide, wherein the conjugate group is cleaved by endogenousnucleases within the body.

Lipid moieties have been used in nucleic acid therapies in a variety ofmethods. In certain such methods, the nucleic acid, such as anoligomeric compound, is introduced into preformed liposomes orlipoplexes made of mixtures of cationic lipids and neutral lipids. Incertain methods, DNA complexes with mono- or poly-cationic lipids areformed without the presence of a neutral lipid. In certain embodiments,a lipid moiety is selected to increase distribution of a pharmaceuticalagent to a particular cell or tissue. In certain embodiments, a lipidmoiety is selected to increase distribution of a pharmaceutical agent tofat tissue. In certain embodiments, a lipid moiety is selected toincrease distribution of a pharmaceutical agent to muscle tissue.

In certain embodiments, pharmaceutical compositions comprise a deliverysystem. Examples of delivery systems include, but are not limited to,liposomes and emulsions. Certain delivery systems are useful forpreparing certain pharmaceutical compositions including those comprisinghydrophobic compounds. In certain embodiments, certain organic solventssuch as dimethylsulfoxide are used.

In certain embodiments, pharmaceutical compositions comprise one or moretissue-specific delivery molecules designed to deliver the one or morepharmaceutical agents comprising an oligomeric compound provided hereinto specific tissues or cell types. For example, in certain embodiments,pharmaceutical compositions include liposomes coated with atissue-specific antibody.

In certain embodiments, pharmaceutical compositions comprise aco-solvent system. Certain of such co-solvent systems comprise, forexample, benzyl alcohol, a nonpolar surfactant, a water-miscible organicpolymer, and an aqueous phase. In certain embodiments, such co-solventsystems are used for hydrophobic compounds. A non-limiting example ofsuch a co-solvent system is the VPD co-solvent system, which is asolution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v ofthe nonpolar surfactant Polysorbate 80™ and 65% w/v polyethylene glycol300. The proportions of such co-solvent systems may be variedconsiderably without significantly altering their solubility andtoxicity characteristics. Furthermore, the identity of co-solventcomponents may be varied: for example, other surfactants may be usedinstead of Polysorbate 80™; the fraction size of polyethylene glycol maybe varied; other biocompatible polymers may replace polyethylene glycol,e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides maysubstitute for dextrose.

In certain embodiments, pharmaceutical compositions are prepared fororal administration. In certain embodiments, pharmaceutical compositionsare prepared for buccal administration. In certain embodiments, apharmaceutical composition is prepared for administration by injection(e.g., intravenous, subcutaneous, intramuscular, intrathecal (IT),intracerebroventricular (ICV), intraneural, perineural, etc.). Incertain of such embodiments, a pharmaceutical composition comprises acarrier and is formulated in aqueous solution, such as water orphysiologically compatible buffers such as Hanks

solution, Ringer

solution, or physiological saline buffer. In certain embodiments, otheringredients are included (e.g., ingredients that aid in solubility orserve as preservatives). In certain embodiments, injectable suspensionsare prepared using appropriate liquid carriers, suspending agents andthe like. Certain pharmaceutical compositions for injection arepresented in unit dosage form, e.g., in ampoules or in multi-dosecontainers. Certain pharmaceutical compositions for injection aresuspensions, solutions or emulsions in oily or aqueous vehicles, and maycontain formulatory agents such as suspending, stabilizing and/ordispersing agents. Certain solvents suitable for use in pharmaceuticalcompositions for injection include, but are not limited to, lipophilicsolvents and fatty oils, such as sesame oil, synthetic fatty acidesters, such as ethyl oleate or triglycerides, and liposomes.

Under certain conditions, certain compounds disclosed herein act asacids. Although such compounds may be drawn or described in protonated(free acid) form, or ionized and in association with a cation (salt)form, aqueous solutions of such compounds exist in equilibrium amongsuch forms. For example, a phosphate linkage of an oligonucleotide inaqueous solution exists in equilibrium among free acid, anion and saltforms. Unless otherwise indicated, compounds described herein areintended to include all such forms. Moreover, certain oligonucleotideshave several such linkages, each of which is in equilibrium. Thus,oligonucleotides in solution exist in an ensemble of forms at multiplepositions all at equilibrium. The term “oligonucleotide” is intended toinclude all such forms. Drawn structures necessarily depict a singleform. Nevertheless, unless otherwise indicated, such drawings arelikewise intended to include corresponding forms. Herein, a structuredepicting the free acid of a compound followed by the term “or saltthereof” expressly includes all such forms that may be fully orpartially protonated/de-protonated/in association with a cation. Incertain instances, one or more specific cation is identified.

In certain embodiments, modified oligonucleotides or oligomericcompounds are in aqueous solution with sodium. In certain embodiments,modified oligonucleotides or oligomeric compounds are in aqueoussolution with potassium. In certain embodiments, modifiedoligonucleotides or oligomeric compounds are in PBS. In certainembodiments, modified oligonucleotides or oligomeric compounds are inwater. In certain such embodiments, the pH of the solution is adjustedwith NaOH and/or HCl to achieve a desired pH.

Herein, certain specific doses are described. A dose may be in the formof a dosage unit. For clarity, a dose (or dosage unit) of a modifiedoligonucleotide or an oligomeric compound in milligrams indicates themass of the free acid form of the modified oligonucleotide or oligomericcompound. As described above, in aqueous solution, the free acid is inequilibrium with anionic and salt forms. However, for the purpose ofcalculating dose, it is assumed that the modified oligonucleotide oroligomeric compound exists as a solvent-free, sodium-acetate free,anhydrous, free acid. For example, where a modified oligonucleotide oran oligomeric compound is in solution comprising sodium (e.g., saline),the modified oligonucleotide or oligomeric compound may be partially orfully de-protonated and in association with Na+ ions. However, the massof the protons are nevertheless counted toward the weight of the dose,and the mass of the Na+ ions are not counted toward the weight of thedose. Thus, for example, a dose, or dosage unit, of 10 mg of CompoundNo. 1362458, equals the number of fully protonated molecules that weighs10 mg. This would be equivalent to 10.47 mg of solvent-free, sodiumacetate-free, anhydrous sodiated Compound No. 1362458. When anoligomeric compound comprises a conjugate group, the mass of theconjugate group is included in calculating the dose of such oligomericcompound. If the conjugate group also has an acid, the conjugate groupis likewise assumed to be fully protonated for the purpose ofcalculating dose.

VI. Certain Compositions

1. Compound No. 1166998

In certain embodiments, Compound No. 1166998 is characterized as a6-10-4 MOE gapmer having a sequence (from 5′ to 3′) ofCAGTATTACCTCTACTAGTC (SEQ ID NO: 20), wherein each of nucleosides 1-6and 17-20 (from 5′ to 3′) are 2′-β-D-MOE nucleosides and each ofnucleosides 7-16 are 2′-β-D-deoxynucleosides, wherein theinternucleoside linkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 5to 6, 6 to 7, and 17 to 18 are phosphodiester internucleoside linkagesand the internucleoside linkages between nucleosides 1 to 2, 7 to 8, 8to 9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to16, 16 to 17, 18 to 19, and 19 to 20 are phosphorothioateinternucleoside linkages, and wherein each cytosine is a5-methylcytosine.

In certain embodiments, Compound No. 1166998 is represented by thefollowing chemical notation:^(m)C_(es)A_(eo)G_(eo)T_(eo)A_(eo)T_(eo)T_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds) ^(m)C_(ds)T_(ds)A_(ds)^(m)C_(ds)T_(ds)A_(eo)G_(es)T_(es) ^(m)C_(e) (SEQ ID NO: 20), wherein:

A=an adenine nucleobase,

^(m)C=a 5-methylcytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-β-D-MOE sugar moiety,

d=a 2′-β-D-deoxyribosyl sugar moiety,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No. 1166998 is represented by thefollowing chemical structure:

Structure 1. Compound No 116699

In certain embodiments, the sodium salt of Compound No. 1166998 isrepresented by the following chemical structure:

Structure 2. The Sodium Salt of Compound No. 1166998

2. Compound No. 1166985

In certain embodiments, Compound No. 1166985 is characterized as a6-10-4 MOE gapmer having a sequence (from 5′ to 3′) ofCACATTCACTAATATTTAAC (SEQ ID NO: 21), wherein each of nucleosides 1-6and 17-20 (from 5′ to 3′) are 2′-β-D-MOE nucleosides and each ofnucleosides 7-16 are 2′-β-D-deoxynucleosides, wherein theinternucleoside linkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 5to 6, 6 to 7, and 17 to 18 are phosphodiester internucleoside linkagesand the internucleoside linkages between nucleosides 1 to 2, 7 to 8, 8to 9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to16, 16 to 17, 18 to 19, and 19 to 20 are phosphorothioateinternucleoside linkages, and wherein each cytosine is a5-methylcytosine.

In certain embodiments, Compound No. 1166985 is represented by thefollowing chemical notation: ^(m)C_(es)A_(eo)^(m)C_(eo)A_(eo)T_(eo)T_(eo) ^(m)C_(ds)A_(ds)^(m)C_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)T_(ds)T_(ds)T_(eo)A_(es)A_(es)^(m)C_(e) (SEQ ID NO: 21), wherein:

A=an adenine nucleobase,

^(m)C=a 5-methylcytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-β-D-MOE sugar moiety,

d=a 2′-β-D-deoxyribosyl sugar moiety,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No. 1166985 is represented by thefollowing chemical structure:

Structure 3. Compound No. 1166985

In certain embodiments, the sodium salt of Compound No. 1166985 isrepresented by the following chemical structure:

Structure 4. The Sodium Salt of Compound No. 1166985

3. Compound No. 1166954

In certain embodiments, Compound No. 1166954 is characterized as a5-10-5 MOE gapmer having a sequence (from 5′ to 3′) ofCCAGTGTCTTCACTTTGCTC (SEQ ID NO: 825), wherein each of nucleosides 1-5and 16-20 (from 5′ to 3′) are 2′-β-D-MOE nucleosides and each ofnucleosides 6-15 are 2′-β-D-deoxynucleosides, wherein theinternucleoside linkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 5to 6, 16 to 17, and 17 to 18 are phosphodiester internucleoside linkagesand the internucleoside linkages between nucleosides 1 to 2, 6 to 7, 7to 8, 8 to 9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15,15 to 16, 18 to 19, and 19 to 20 are phosphorothioate internucleosidelinkages, and wherein each cytosine is a 5-methylcytosine.

In certain embodiments, Compound No. 1166954 is represented by thefollowing chemical notation: ^(m)C_(es)^(m)C_(eo)A_(eo)G_(eo)T_(eo)G_(ds)T_(ds) ^(m)C_(ds)T_(ds)T_(ds)^(m)C_(ds)A_(ds) ^(m)C_(ds)T_(ds)T_(ds)T_(eo)G_(eo) ^(m)C_(es)T_(es)^(m)C_(e) (SEQ ID NO: 825), wherein:

A=an adenine nucleobase,

^(m)C=a 5-methylcytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-β-D-MOE sugar moiety,

d=a 2′-β-D-deoxyribosyl sugar moiety,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No. 1166954 is represented by thefollowing chemical structure:

Structure 5: Compound No. 1166954

In certain embodiments, the sodium salt of Compound No. 1166954 isrepresented by the following chemical structure:

Structure 6: The Sodium Salt of Compound No. 1166954

4. Compound No. 1072813

In certain embodiments, Compound No. 1072813 is characterized as a5-10-5 MOE gapmer having a sequence (from 5′ to 3′) ofGCAACAGTTTCCATAACAAC (SEQ ID NO: 1499), wherein each of nucleosides 1-5and 16-20 (from 5′ to 3′) are 2′-β-D-MOE nucleosides and each ofnucleosides 6-15 are 2′-β-D-deoxynucleosides, wherein theinternucleoside linkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 16to 17, and 17 to 18 are phosphodiester internucleoside linkages and theinternucleoside linkages between nucleosides 1 to 2, 5 to 6, 6 to 7, 7to 8, 8 to 9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15,15 to 16, 18 to 19, and 19 to 20 are phosphorothioate internucleosidelinkages, and wherein each cytosine is a 5-methylcytosine.

In certain embodiments, Compound No. 1072813 is represented by thefollowing chemical notation: G_(es) ^(m)C_(eo)A_(eo)A_(eo)^(m)C_(es)A_(ds)G_(ds)T_(ds)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)A_(ds)T_(ds)A_(ds)A_(eo) ^(m)C_(eo)A_(es)A_(es) ^(m)C_(e) (SEQID NO: 1499), wherein:

A=an adenine nucleobase,

^(m)C=a 5-methylcytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-β-D-MOE sugar moiety,

d=a 2′-β-D-deoxyribosyl sugar moiety,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No. 1072813 is represented by thefollowing chemical structure:

Structure 7: Compound No. 1072813

In certain embodiments, the sodium salt of Compound No. 1072813 isrepresented by the following chemical structure:

Structure 8: The Sodium Salt of Compound No. 1072813

5. Compound No. 1199983

In certain embodiments, Compound No. 1199983 is characterized as a5-10-5 MOE gapmer having a sequence (from 5′ to 3′) ofTGGTCCTAAATATTCTAGTC (SEQ ID NO: 2170), wherein each of nucleosides 1-5and 16-20 (from 5′ to 3′) are 2′-β-D-MOE nucleosides and each ofnucleosides 6-15 are 2′-β-D-deoxynucleosides, wherein theinternucleoside linkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 5to 6, 16 to 17, and 17 to 18 are phosphodiester internucleoside linkagesand the internucleoside linkages between nucleosides 1 to 2, 6 to 7, 7to 8, 8 to 9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15,15 to 16, 18 to 19, and 19 to 20 are phosphorothioate internucleosidelinkages, and wherein each cytosine is a 5-methylcytosine.

In certain embodiments, Compound No. 1199983 is represented by thefollowing chemical notation: T_(es)G_(eo)G_(eo)T_(eo) ^(m)C_(eo)^(m)C_(ds)T_(ds)A_(ds)A_(ds)A_(ds)T_(ds)A_(ds)T_(ds)T_(ds)^(m)C_(ds)T_(eo)A_(eo)G_(es)T_(es) ^(m)C_(e) (SEQ ID NO: 2170), wherein:

A=an adenine nucleobase,

^(m)C=a 5-methylcytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-β-D-MOE sugar moiety,

d=a 2′-β-D-deoxyribosyl sugar moiety,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No. 1199983 is represented by thefollowing chemical structure:

Structure 9: Compound No. 1199983

In certain embodiments, the sodium salt of Compound No. 1199983 isrepresented by the following chemical structure:

Structure 10: The Sodium Salt of Compound No. 1199983

6. Compound No. 1166721

In certain embodiments, Compound No. 1166721 is characterized as a 5-8-5MOE gapmer having a sequence (from 5′ to 3′) of TGGTCCTAAATATTCTAGTC(SEQ ID NO: 2818), wherein each of nucleosides 1-5 and 14-18 (from 5′ to3′) are 2′-β-D-MOE nucleosides and each of nucleosides 6-15 are2′-β-D-deoxynucleosides, wherein the internucleoside linkages betweennucleosides 2 to 3, 3 to 4, 4 to 5, 14 to 15 and 15 to 16 arephosphodiester internucleoside linkages and the internucleoside linkagesbetween nucleosides 1 to 2, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9 to 10, 10to 11, 11 to 12, 12 to 13, 13 to 14, 16 to 17, and 17 to 18 arephosphorothioate internucleoside linkages, and wherein each cytosine isa 5-methylcytosine.

In certain embodiments, Compound No. 1166721 is represented by thefollowing chemical notation: T_(es)G_(eo)G_(eo)^(m)C_(eo)A_(es)G_(ds)T_(ds)A_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)T_(eo) ^(m)C_(eo)T_(es)A_(es) ^(m)C_(e) (SEQ ID NO: 2818),wherein:

A=an adenine nucleobase,

^(m)C=a 5-methylcytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-β-D-MOE sugar moiety,

d=a 2′-β-D-deoxyribosyl sugar moiety,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No. 1166721 is represented by thefollowing chemical structure:

Structure 11: Compound No. 1166721

In certain embodiments, the sodium salt of Compound No. 1166721 isrepresented by the following chemical structure:

Structure 12: The sodium salt of Compound No. 1166721

VIII. Certain Hotspot Regions

In certain embodiments, nucleobases in the ranges specified belowcomprise a hotspot region of GFAP nucleic acid. In certain embodiments,modified oligonucleotides that are complementary to a hotspot region ofGFAP nucleic acid achieve an average of more than 50% reduction of GFAPRNA in vitro in the standard cell assay. In certain embodiments,modified oligonucleotides that are complementary to a hotspot region ofGFAP nucleic acid achieve an average of 75% or greater reduction of GFAPRNA in vivo in the standard in vivo assay.

1. Nucleobases 9324-9348 of SEQ ID NO: 2

In certain embodiments, nucleobases 9324-9348 of SEQ ID NO: 2 comprise ahotspot region. In certain embodiments, modified oligonucleotides arecomplementary to a portion of nucleobases 9324-9348 of SEQ ID NO: 2. Incertain embodiments, modified oligonucleotides are 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are 18nucleobases in length. In certain embodiments, modified oligonucleotidesare gapmers. In certain embodiments, the gapmers are MOE gapmers. Incertain embodiments, the internucleoside linkages of the modifiedoligonucleotides are phosphorothioate internucleoside linkages andphosphodiester internucleoside linkages. In certain embodiments, thephosphodiester (“o”) and phosphorothioate (“s”) internucleoside linkagesare arranged in order from 5′ to 3′: In certain embodiments, modifiednucleotides have an internucleoside linkage motif ofsooosssssssssssooss, sooooossssssssssoss, soooossssssssssooss,sooosssssssssooss, or sooossssssssssoooss, wherein each “s” represents aphosphorothioate internucleoside linkage and each “o” represents aphosphodiester internucleoside linkage.

The nucleobase sequences of SEQ ID Nos: 21, 1177, 2321, 2398, 2808-2809,2840-2842, and 2853-2854 are complementary to a portion of nucleobases9324-9348 of SEQ ID NO: 2.

The nucleobase sequence of Compound Nos.: 1048181-1048182,1104071-1104072, 1166746-1166748, 1166803-1166808, 1166894-1166899,1166985-1166990, 1174016, and 1174018 are complementary to a portion ofnucleobases 9324-9348 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary to aportion of nucleobases 9324-9348 of SEQ ID NO: 2 achieve at least 7%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 9324-9348 of SEQ ID NO: 2 achieve an average of 42%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 9324-9348 of SEQ ID NO: 2 achieve an average of 81%reduction of GFAP RNA in vivo in the standard in vivo assay.

2. Nucleobases 9459-9480 of SEQ ID NO: 2

In certain embodiments, nucleobases 9459-9480 of SEQ ID NO: 2 comprise ahotspot region. In certain embodiments, modified oligonucleotides arecomplementary to a portion of nucleobases 9459-9480 of SEQ ID NO: 2. Incertain embodiments, modified oligonucleotides are 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are 18nucleobases in length. In certain embodiments, modified oligonucleotidesare gapmers. In certain embodiments, the gapmers are MOE gapmers. Incertain embodiments, the internucleoside linkages of the modifiedoligonucleotides are phosphorothioate internucleoside linkages andphosphodiester internucleoside linkages. In certain embodiments, thephosphodiester (“o”) and phosphorothioate (“s”) internucleoside linkagesare arranged in order from 5′ to 3′: In certain embodiments, modifiednucleotides have an internucleoside linkage motif of sooosssssssssssoossor soooossssssssssooss, wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage.

The nucleobase sequences of SEQ ID Nos: 555, 2093, 2170, and 2813 arecomplementary to a portion of nucleobases 9459-9480 of SEQ ID NO: 2.

The nucleobase sequence of Compound Nos.: 1048190, 1104116-1104117, and1199982-1199984 are complementary to a portion of nucleobases 9459-9480of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary to aportion of nucleobases 9459-9480 of SEQ ID NO: 2 achieve at least 26%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 9459-9480 of SEQ ID NO: 2 achieve an average of 42%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 9459-9480 of SEQ ID NO: 2 achieve an average of 91%reduction of GFAP RNA in vivo in the standard in vivo assay.

3. Nucleobases 9530-9580 of SEQ ID NO: 2

In certain embodiments, nucleobases 9530-9580 of SEQ ID NO: 2 comprise ahotspot region. In certain embodiments, modified oligonucleotides arecomplementary to a portion of nucleobases 9530-9580 of SEQ ID NO: 2. Incertain embodiments, modified oligonucleotides are 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are 18nucleobases in length. In certain embodiments, modified oligonucleotidesare gapmers. In certain embodiments, the gapmers are MOE gapmers. Incertain embodiments, the internucleoside linkages of the modifiedoligonucleotides are phosphorothioate internucleoside linkages andphosphodiester internucleoside linkages. In certain embodiments, thephosphodiester (“o”) and phosphorothioate (“s”) internucleoside linkagesare arranged in order from 5′ to 3′: In certain embodiments, modifiednucleotides have an internucleoside linkage motif ofsooosssssssssssooss, sooooossssssssssoss, soooossssssssssooss,sooosssssssssooss, or sooossssssssssoooss, wherein each “s” represents aphosphorothioate internucleoside linkage and each “o” represents aphosphodiester internucleoside linkage.

The nucleobase sequences of SEQ ID Nos.: 20, 88, 166, 1331, 1408, 1485,1637, 1713, 1714, 1789, 1790, 1637, 1638, 1865, 1866, 1941, 2018, 2095,2172, 2249, 2326, 2403, 2480, 2557, 2633, 2709, 2785, 2816-2818, 2859,2861, and 2886-2887 are complementary to a portion of nucleobases9530-9580 of SEQ ID NO: 2.

The nucleobase sequence of Compound Nos.: 1048200-1048201,1073062-1073064, 1104142-1104161, 1166719-1166721, 1166816-1166823,1166826, 1166907-1166920, 1166998-1167011, 1174024, 1174026, and1174029-1174030 are complementary to a portion of nucleobases 9530-9580of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary to aportion of nucleobases 9530-9580 of SEQ ID NO: 2 achieve at least 27%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 9530-9580 of SEQ ID NO: 2 achieve an average of 52%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 9530-9580 of SEQ ID NO: 2 achieve an average of 82%reduction of GFAP RNA in vivo in the standard in vivo assay.

4. Nucleobases 12006-12038 of SEQ ID NO: 2

In certain embodiments, nucleobases 12006-12038 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to a portion of nucleobases 12006-12038 of SEQ ID NO: 2.In certain embodiments, modified oligonucleotides are 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are 18nucleobases in length. In certain embodiments, modified oligonucleotidesare gapmers. In certain embodiments, the gapmers are MOE gapmers. Incertain embodiments, the internucleoside linkages of the modifiedoligonucleotides are phosphorothioate internucleoside linkages andphosphodiester internucleoside linkages. In certain embodiments, thephosphodiester (“o”) and phosphorothioate (“s”) internucleoside linkagesare arranged in order from 5′ to 3′: In certain embodiments, modifiednucleotides have an internucleoside linkage motif ofsooosssssssssssooss, wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage.

The nucleobase sequences of SEQ ID Nos.: 815, 893, 971, 1049, 1269,1270, 1346, 1423, 1499, 1500, 1660, 1736, 2655, and 2731 arecomplementary to a portion of nucleobases 12006-12038 of SEQ ID NO: 2.

The nucleobase sequence of Compound Nos.: 1047362-1047365,1072813-1072818, and 1103276-1103279 are complementary to a portion ofnucleobases 12006-12038 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary to aportion of nucleobases 12006-12038 of SEQ ID NO: 2 achieve at least 29%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 12006-12038 of SEQ ID NO: 2 achieve an average of 52%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 12006-12038 of SEQ ID NO: 2 achieve an average of 82%reduction of GFAP RNA in vivo in the standard in vivo assay.

5. Nucleobases 13038-13058 of SEQ ID NO: 2

In certain embodiments, nucleobases 13038-13058 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to a portion of nucleobases 13038-13058 of SEQ ID NO: 2.In certain embodiments, modified oligonucleotides are 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are 18nucleobases in length. In certain embodiments, modified oligonucleotidesare gapmers. In certain embodiments, the gapmers are MOE gapmers. Incertain embodiments, the internucleoside linkages of the modifiedoligonucleotides are phosphorothioate internucleoside linkages andphosphodiester internucleoside linkages. In certain embodiments, thephosphodiester (“o”) and phosphorothioate (“s”) internucleoside linkagesare arranged in order from 5′ to 3′: In certain embodiments, modifiednucleotides have an internucleoside linkage motif ofsooosssssssssssooss, sooooossssssssssoss, or soooossssssssssooss,wherein each “s” represents a phosphorothioate internucleoside linkageand each “o” represents a phosphodiester internucleoside linkage.

The nucleobase sequences of SEQ ID Nos.: 825 and 1973 are complementaryto a portion of nucleobases 13038-13058 of SEQ ID NO: 2.

The nucleobase sequence of Compound Nos.: 1047522, 1166954, and 1167046are complementary to a portion of nucleobases 13038-13058 of SEQ ID NO:2.

In certain embodiments, modified oligonucleotides complementary to aportion of nucleobases 13038-13058 of SEQ ID NO: 2 achieve at least 27%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 13038-13058 of SEQ ID NO: 2 achieve an average of 41%reduction of GFAP RNA in vitro in the standard cell assay. In certainembodiments, modified oligonucleotides complementary to a portion ofnucleobases 13038-13058 of SEQ ID NO: 2 achieve an average of 84%reduction of GFAP RNA in vivo in the standard in vivo assay.

6. Additional Hotspot Regions

In certain embodiments, the ranges described in the Table below comprisehotspot regions. Each hotspot region begins with the nucleobase of SEQID NO: 2 identified in the “Start Site SEQ ID NO: 2” column and endswith the nucleobase of SEQ ID NO: 2 identified in the “Stop Site SEQ IDNO: 2” column. In certain embodiments, modified oligonucleotides arecomplementary within any of the hotspot regions 1-14, as defined in thetable below. In certain embodiments, modified oligonucleotides are 18nucleobases in length. In certain embodiments, modified oligonucleotidesare 20 nucleobases in length. In certain embodiments, modifiedoligonucleotides are gapmers. In certain embodiments, modifiedoligonucleotides are 5-8-5, 5-10-3, 4-10-6, 6-10-4, or 5-10-5 MOEgapmers.

The nucleobase sequence of compounds listed in the “Compound No. inrange” column in the table below are complementary to SEQ ID NO: 2within the specified hotspot region. The nucleobase sequence of theoligonucleotides listed in the “SEQ ID NO: in range” column in the tablebelow are complementary to the target sequence, SEQ ID NO: 2, within thespecified hotspot region.

In certain embodiments, modified oligonucleotides complementary tonucleobases within the hotspot region achieve at least “Sin.% Red. invitro” (minimum % reduction, relative to untreated control cells) ofGFAP RNA in vitro in the standard cell assay, as indicated in the tablebelow. In certain embodiments, modified oligonucleotides complementaryto nucleobases within the hotspot region achieve an average of “Avg.%Red. in vitro” (average % reduction, relative to untreated controlcells) of GFAP RNA in vitro in the standard cell assay, as indicated inthe table below. In certain embodiments, modified oligonucleotidescomplementary to nucleobases within the hotspot region achieve a maximumof “Max. % Red. in vitro” (maximum % reduction, relative to untreatedcontrol cells) of GFAP RNA in vitro in the standard cell assay, asindicated in the table below. In certain embodiments, modifiedoligonucleotides complementary to nucleobases within the hotspot regionachieve an average of “Avg. % Red. in vivo” (average % reduction,relative to PBS-treated animals) of GFAP RNA in vivo in the standard invivo assay, as indicated in the table below.

TABLE 1 GFAP Hotspots Start Start Site Site Min. Max. Avg. Avg. HotspotSEQ ID SEQ ID % Red. % Red. % Red. % Red. Compound No. SEQ ID NOs: IDNO: 2 NO: 2 in vitro In vitro in vitro in vivo in Range in Range 1 93249348 7 78 42 81 1048181-1048182, 21, 1177, 2321, 2398, 1104071-1104072,2808-2809, 2840-2842, 1166746-1166748, 2853-2854 1166803-1166808,1166894-1166899, 1166985-1166990, 1174016, 1174018 2 9459 9480 26 65 4291 1048190, 555, 2093, 2170, 1104116-1104117, 2813 1199982-1199984 39530 9580 18 82 52 82 1048199-1048201, 20, 88, 166, 1331,1073062-1073064, 1408, 1485, 1637, 1104142-1104161, 1713, 1714, 1789,1166719-1166721, 1790, 1637, 1638, 1166816-1166823, 1865, 1866, 1941,1166826, 2018, 2095, 2172, 1166907-1166920, 2249, 2326, 2403,1166998-1167011, 2480, 2557, 2633, 1174024, 1174026, 2709, 2785,1174029-1174030 2816-2818, 2859, 2861, 2886-2887 4 12006 12038 29 75 5282 1047362-1047365, 815, 893, 971, 1049, 1072813-1072818, 1269, 1270,1346, 1103276-1103279 1423, 1499, 1500, 1660, 1736, 2655, 2731 5 1303813058 27 56 41 84 1047522, 1166954, 825, 1973 1167046 6 8530 8557 14 7952 89 1047706-1047708, 213, 291, 369, 1601, 1103567-1103571 1753, 1829,1905, 1982 7 8731 8754 44 64 53 88 1047733-1047735, 1072, 1149, 1227,1103591, 1103592, 2291, 2368 1174050, 1174051, 1174056, 1174058,1174062,1174063 8 8749 8807 25 92 52 75 1047601-1047610, 51, 129, 207,752, 1072854-1072868, 830, 908, 986, 1064, 1103462-1103472, 1141, 1219,1279, 1166738-1166740, 1280-1282, 1356-1359, 1166742, 1166744,1433-1436, 1510-1512, 1166793-1166795, 1595, 1671, 1747,1166798-1166800, 2206, 2283, 2360, 1166885-1166890, 2437, 2514, 2590,1166975, 1166982, 2666, 2742, 2835-2837, 1174012-1174013 2839,2850-2851, 2866 9 9511 9536 32 90 55 86 1048197, 1073060, 1100, 1484,1864, 1104129-1104133, 1940, 2017, 2094, 1166749-1166751, 2171,2819-2821, 1166809-1166810, 2856 1166812-1166813, 1166900-1166903,1166991-1166994, 1174020 10 9565 9602 20 75 53 82 1048202-1048204, 244,322, 400, 1562, 1073065, 1639, 1715, 1791, 1104165-1104179, 1867, 1943,2020, 1166757-1166760, 2173, 2250, 2327, 1166831, 1166835, 2404, 2481,2558, 1166926-1166929, 2634, 2710, 2786, 1167012, 1167017, 2873-2876,2888, 1167018, 1174031, 2891 1174034 11 11155 11184 37 82 69 851073093-1073095, 1339, 1569, 2028, 1104307-1104312, 2105, 2182, 2259,1167024-1167027, 2336, 2413, 1166761-1166763, 2822-2824, 28631166842-1166843, 1166845-1166847, 1166933-1166934, 1167024-1167027,1174037 12 12044 12067 49 67 58 85 1047372-1047374, 348, 426, 504, 1425,1072824-1072825, 1502 1166852, 1167033-1167036 13 12080 12108 45 88 6879 1047384-1047388, 37, 115, 193, 271, 1072834-1072835, 349, 1274, 1351,1103284-1103285, 2041, 2118 1166948, 1167040 14 13333 13367 53 93 75 891047579-104592, 50, 283, 361, 439, 1072849-1072850, 517, 595, 673, 751,1166775-1166787, 829, 907, 985, 1063, 1166867, 1140, 1218, 1278,1166869-1166875, 1508, 2825-2833, 1166877-1166878, 2843-28461167050-1167051, 1167053-1167060Nonlimiting Disclosure and Incorporation by Reference

Each of the literature and patent publications listed herein isincorporated by reference in its entirety.

While certain compounds, compositions and methods described herein havebeen described with specificity in accordance with certain embodiments,the following examples serve only to illustrate the compounds describedherein and are not intended to limit the same. Each of the references,GenBank accession numbers, and the like recited in the presentapplication is incorporated herein by reference in its entirety.

Although the sequence listing accompanying this filing identifies eachsequence as either “RNA” or “DNA” as required, in reality, thosesequences may be modified with any combination of chemicalmodifications. One of skill in the art will readily appreciate that suchdesignation as “RNA” or “DNA” to describe modified oligonucleotides is,in certain instances, arbitrary. For example, an oligonucleotidecomprising a nucleoside comprising a 2′-OH sugar moiety and a thyminebase could be described as a DNA having a modified sugar moiety (2′-OHin place of one 2′-H of DNA) or as an RNA having a modified base(thymine (methylated uracil) in place of a uracil of RNA). Accordingly,nucleic acid sequences provided herein, including, but not limited tothose in the sequence listing, are intended to encompass nucleic acidscontaining any combination of natural or modified RNA and/or DNA,including, but not limited to such nucleic acids having modifiednucleobases. By way of further example and without limitation, anoligomeric compound having the nucleobase sequence “ATCGATCG”encompasses any oligomeric compounds having such nucleobase sequence,whether modified or unmodified, including, but not limited to, suchcompounds comprising RNA bases, such as those having sequence “AUCGAUCG”and those having some DNA bases and some RNA bases such as “AUCGATCG”and oligomeric compounds having other modified nucleobases, such as“AT^(m)CGAUCG,” wherein ^(m)C indicates a cytosine base comprising amethyl group at the 5-position.

Certain compounds described herein (e.g., modified oligonucleotides)have one or more asymmetric center and thus give rise to enantiomers,diastereomers, and other stereoisomeric configurations that may bedefined, in terms of absolute stereochemistry, as (R) or (S), as α or βsuch as for sugar anomers, or as (D) or (L), such as for amino acids,etc. Compounds provided herein that are drawn or described as havingcertain stereoisomeric configurations include only the indicatedcompounds. Compounds provided herein that are drawn or described withundefined stereochemistry include all such possible isomers, includingtheir stereorandom and optically pure forms, unless specified otherwise.Likewise, all cis- and trans-isomers and tautomeric forms of thecompounds herein are also included unless otherwise indicated.Oligomeric compounds described herein include chirally pure or enrichedmixtures as well as racemic mixtures. For example, oligomeric compoundshaving a plurality of phosphorothioate internucleoside linkages includesuch compounds in which chirality of the phosphorothioateinternucleoside linkages is controlled or is random. Unless otherwiseindicated, compounds described herein are intended to includecorresponding salt forms.

The compounds described herein include variations in which one or moreatoms are replaced with a non-radioactive isotope or radioactive isotopeof the indicated element. For example, compounds herein that comprisehydrogen atoms encompass all possible deuterium substitutions for eachof the ¹H hydrogen atoms. Isotopic substitutions encompassed by thecompounds herein include but are not limited to: ²H or ³H in place of¹H, ¹³C or ¹⁴C in place of ¹²C, ¹⁵N in place of ¹⁴N, ¹⁷O or ¹⁸O in placeof ¹⁶O and ³³S, ³⁴S, ³⁵S, or ³⁶S in place of ³²S. In certainembodiments, non-radioactive isotopic substitutions may impart newproperties on the oligomeric compound that are beneficial for use as atherapeutic or research tool. In certain embodiments, radioactiveisotopic substitutions may make the compound suitable for research ordiagnostic purposes such as imaging.

EXAMPLES

The following examples illustrate certain embodiments of the presentdisclosure and are not limiting. Moreover, where specific embodimentsare provided, the inventors have contemplated generic application ofthose specific embodiments.

Example 1: Effect of 5-10-5 MOE Gapmer Modified Oligonucleotides onHuman GFAP RNA In Vitro, Single Dose

Modified oligonucleotides complementary to human GFAP nucleic acid weredesigned and tested for their single dose effects on GFAP RNA in vitro.The modified oligonucleotides were tested in a series of experimentsthat had similar culture conditions.

The modified oligonucleotides in the tables below are 5-10-5 MOE gapmerswith mixed PO/PS internucleoside linkages. The gapmers are 20nucleosides in length, wherein the central gap segment consists of ten2′-β-D-deoxynucleosides and the 5′ and 3′ wing segments each consists offive 2′-MOE modified nucleosides. The sugar motif for the gapmers is(from 5′ to 3′): eeeeeddddddddddeeeee; wherein ‘d’ represents a2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-MOE sugarmoiety. The internucleoside linkage motif for the gapmers is (from 5′ to3′): sooosssssssssssooss; wherein each ‘o’ represents a phosphodiesterinternucleoside linkage and each ‘s’ represents a phosphorothioateinternucleoside linkage. Each cytosine residue is a 5-methylcytosine.

“Start site” indicates the 5′-most nucleoside to which the modifiedoligonucleotide is complementary in the target nucleic acid sequence.“Stop site” indicates the 3′-most nucleoside to which the modifiedoligonucleotide is complementary in the target nucleic acid sequence.Each modified oligonucleotide listed in the Tables below is 100%complementary to SEQ ID NO: 1 (GENBANK Accession No. NM_002055.4), orSEQ ID NO: 2 (GENBANK Accession No. NC_000017.11 truncated fromnucleotides 44903001 to 44919000). ‘N/A’ indicates that the modifiedoligonucleotide is not 100% complementary to that particular targetnucleic acid sequence.

Cultured U251 cells were treated with modified oligonucleotide at aconcentration of 4,000 nM using free uptake at a density of 10,000 cellsper well. After a treatment period of approximately 48 hours, total RNAwas isolated from the cells and GFAP RNA levels were measured byquantitative real-time RTPCR. GFAP RNA levels were measured by humanGFAP primer probe set RTS37485 (forward sequence CTGGAGGTTGAGAGGGACA,designated herein as SEQ ID NO: 11; reverse sequenceGCTTCATCTGCTTCCTGTCT, designated herein as SEQ ID NO: 12; probe sequenceCTGGAGCTTCTGCCTCACAGTGG, designated herein as SEQ ID NO: 13). GFAP RNAlevels were normalized to total RNA content, as measured by RIBOGREEN®.Reduction of GFAP RNA is presented in the tables below as percent GFAPRNA amount relative to untreated control cells. Each table representsresults from an individual assay plate. The values marked with anasterisk (*) indicate that the modified oligonucleotide is complementaryto the amplicon region of the primer probe set. Additional assays may beused to measure the potency and efficacy of the modifiedoligonucleotides complementary to the amplicon region.

TABLE 2Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 GFAP SEQ Compound Start Stop Start Stop RNA ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047144   17   36  3465  3484CGAGGGCTTTATGAAGGAGT  80 22 1047160  131  150  3579  3598GCCAGGAGCCAGGCCCCCCA  67 23 1047176  248  267  3696  3715GGCCCGGGTCTCCTTGAAGC 109 24 1047192  488  507  3936  3955GTCCTGTGCCAGATTGTCCC   7* 25 1047208  579  598 N/A N/ACATCTGCTTCCTGTCTATAG   6* 26 1047224  649  668  5238  5257CTCAAGAACCGGATCTCCTC 105 27 1047240  813  832  5699  5718ACTCTTCGGCTTCATGCATG  69 28 1047256 1029 1048  7601  7620GCGCCTCCTGATAACTGGCC  80 29 1047272 1251 1270 10865 10884CTTCTGACACAGACTTGGTG  92 30 1047288 1298 1317 10912 10931ATCCCGCATCTCCACGGTCT  90 31 1047304 1351 1370 11640 11659CTGCCTCACATCACATCCTT  87 32 1047320 1416 1435 11705 11724CGGAGCAACTATCCTGCTTC 102 33 1047336 1617 1636 11906 11925AAGCTGCTGGCCATGCCCCT  87 34 1047352 1664 1683 11953 11972TGCCCCCCGCCCTCCTCCCC  83 35 1047368 1736 1755 12025 12044GAGAGAACCTCCATCTCTGG  47 36 1047384 1791 1810 12080 12099TCAGTTTTCCTCCAGCAGCC  55 37 1047400 1854 1873 12143 12162GACAAAACAAGCCTCTGGCC  66 38 1047416 2036 2055 12325 12344CGTCCCCACCCATCTTAGAC  60 39 1047432 2180 2199 12469 12488GTGCTGAGAATCAAGCTCCC  93 40 1047448 2237 2256 12526 12545CCCCCTCTATCCCTCCCAGC  11 41 1047464 2280 2299 12569 12588CTGGGCTTGACCTCTCTGTA  63 42 1047480 2379 2398 12668 12687TGGTCACCCACAACCCCTAC  81 43 1047496 2457 2476 12746 12765CCCTTTCTCTCCTGTTTCAG  79 44 1047512 2486 2505 12775 12794AAGTCATGCCCTGCCCCCAT  45 45 1047528 2780 2799 13069 13088GCACCCGGCCTCCAGGCTGC  76 46 1047544 2861 2880 13150 13169GGCACAGATCCCACCAGTCT 106 47 1047560 2903 2922 13192 13211GAGAGGAGAACCCTGAAGTG  72 48 1047576 3035 3054 13324 13343CCTCAGCGACTAAAGGCAGC  77 49 1047592 3058 3077 13347 13366GCGCAGCATTTGTCTTTATT  47 50 1047608 N/A N/A  8777  8796GCTTTTGAGATATCTTGTGA   8 51 1047624 N/A N/A  9031  9050GTTTAATGTACAGTTACTCT  71 52 1047640 N/A N/A  9070  9089CCAAGGACTCACCACCTTTA  75 53 1047656 N/A N/A  9202  9221AGGGATGAAAGAATAAAGCA  92 54 1047672 N/A N/A  8381  8400CCTGCTGTACTGACCTCGAA  94 55 1047688 N/A N/A  8456  8475ATCCTCAGTCCCAGTCTGGA  54 56 1047704 N/A N/A  8504  8523CTGCAGTGTCACGAAGGCCC  74 57 1047720 N/A N/A  8637  8656TGTCAAGCTCTCACCCAGTT  77 58 1047736 N/A N/A  8737  8756TTGGTGCTTTTGCCCCCTGT  56 59 1047752 N/A N/A  4093  4112GGATAGTGCCCCATCAAGAG 109 60 1047768 N/A N/A  4264  4283AGTCACAAAGCCCAGCCATG  95 61 1047784 N/A N/A  4322  4341GCTTCCAACTCCTCCTTTAT 101 62 1047800 N/A N/A  4359  4378CAGAATCCAATCTCCCTCAT 106 63 1047816 N/A N/A  4405  4424GCTTTGCGCCCAGACCTGCC  68 64 1047832 N/A N/A  4525  4544ATTCCTCTGATCCCAGGTAA  52 65 1047848 N/A N/A  4704  4723CCTTAACTCATTACTAAGGT  69 66 1047864 N/A N/A  4806  4825GAGACCACCCCCACCCAGGA  70 67 1047880 N/A N/A  4868  4887GTCCAGGCTCTTCTGAGGAC  66 68 1047896 N/A N/A  5007  5026GTGGCCATCAATCCTTTCCT 103 69 1047912 N/A N/A  5117  5136CCCCAGGCTCCTTCTCCCCA  74 70 1047928 N/A N/A  5388  5407TGTCTCTACCTGCCAATCTC 100 71 1047944 N/A N/A  5521  5540CTCAGGGTACAGGCCACAGC  90 72 1047960 N/A N/A  5791  5810ACCCTCCTTCCCCCATTCTC  94 73 1047976 N/A N/A  5934  5953AGCTACTACTAATAATAGCA  98 74 1047992 N/A N/A  6023  6042ACTTCGGCTCTCTCATCTGT  75 75 1048008 N/A N/A  6146  6165AACCCAAAACAGACTGGCAG  79 76 1048024 N/A N/A  6281  6300CCCACACTACATATAAGCTC 105 77 1048040 N/A N/A  6329  6348CCTGTCCTGCCTAGCCCAAA  71 78 1048056 N/A N/A  6417  6436GGGCCCTGCCTCTCTGTGCT  81 79 1048072 N/A N/A  6544  6563ATAGCCCTTTCTCCCCTGCC  86 80 1048088 N/A N/A  6959  6978AATCCAGAACCTTCCACACT 114 81 1048104 N/A N/A  7073  7092TGGGACTTTTCCCAACAACT  93 82 1048120 N/A N/A  7412  7431CGCCCTCGACCCAGGTCCTC  57 83 1048136 N/A N/A  7907  7926AGTGACTGCCTGCTATGTGT  94 84 1048152 N/A N/A  7989  8008TTGGAGGGTGACCCAAGTCC  81 85 1048168 N/A N/A  8235  8254ACGCCCTTTTCCTTGCCAGG  74 86 1048184 N/A N/A  9373  9392TAGCTCCCCCCTCCCCCCGC  52 87 1048200 N/A N/A  9534  9553GCAGTATTACCTCTACTAGT  45 88 1048216 N/A N/A  9610  9629CCTGTCCCCTTTCCTCTTTC  74 89 1048232 N/A N/A  9791  9810CCACCAACCAGCCACATGAC  98 90 1048248 N/A N/A  9826  9845ATCAGGAGACCAGAGCTCAA  78 91 1048264 N/A N/A 10622 10641GGCCTGGCTTCATTTCAGCC  66 92 1048280 N/A N/A 10734 10753AGTATGAGAACCTATGCAAC  80 93 1048296 N/A N/A 10793 10812GGGCATGAGCCATCCTCTCC  35 94 1048312 N/A N/A 10942 10961GACTGGGCCCAAATCCCTCC  87 95 1048328 N/A N/A 11063 11082CCTTGCTCTCCTCCAGAATT  67 96 1048344 N/A N/A 11227 11246GCTACTAACTTTAATTCTCT  41 97 1048360 N/A N/A 11321 11340CCTCTTCCCATTCCCCTGGT  62 98 1048376 N/A N/A 11488 11507GGTCTTACTTTTCTTGATAG  72 99

TABLE 3Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 GFAP SEQ Compound Start Stop Start Stop RNA ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047145   18   37  3466  3485GCGAGGGCTTTATGAAGGAG  99 100 1047161  133  152  3581  3600CGGCCAGGAGCCAGGCCCCC  82 101 1047177  267  286  3715  3734TCATCTCTGCCCGCTCACTG  88 102 1047193  490  509  3938  3957AGGTCCTGTGCCAGATTGTC   8* 103 1047209  585  604 N/A N/ATGGCTTCATCTGCTTCCTGT   5* 104 1047225  651  670  5240  5259TCCTCAAGAACCGGATCTCC  37 105 1047241  821  840  5707  5726GCGGTACCACTCTTCGGCTT  77 106 1047257 1051 1070  7623  7642CCCTCTTCCTCCAGCCGCGC  35 107 1047273 1252 1271 10866 10885CCTTCTGACACAGACTTGGT  86 108 1047289 1305 1324 10919 10938CCTCTCCATCCCGCATCTCC  87 109 1047305 1352 1371 11641 11660CCTGCCTCACATCACATCCT  66 110 1047321 1417 1436 11706 11725GCGGAGCAACTATCCTGCTT  96 111 1047337 1631 1650 11920 11939CCTCATTCTAACGCAAGCTG  58 112 1047353 1665 1684 11954 11973GTGCCCCCCGCCCTCCTCCC  46 113 1047369 1739 1758 12028 12047TCCGAGAGAACCTCCATCTC  72 114 1047385 1792 1811 12081 12100CTCAGTTTTCCTCCAGCAGC  50 115 1047401 1876 1895 12165 12184TCCCACCTCATAAAAACCAA  94 116 1047417 2059 2078 12348 12367GGTGACTGCCCCAGGTGGCA  68 117 1047433 2181 2200 12470 12489AGTGCTGAGAATCAAGCTCC  75 118 1047449 2238 2257 12527 12546CCCCCCTCTATCCCTCCCAG  60 119 1047465 2310 2329 12599 12618GTCCCCTCCAGTCTGCACGG  49 120 1047481 2380 2399 12669 12688CTGGTCACCCACAACCCCTA  82 121 1047497 2458 2477 12747 12766CCCCTTTCTCTCCTGTTTCA  29 122 1047513 2491 2510 12780 12799GGACAAAGTCATGCCCTGCC  85 123 1047529 2781 2800 13070 13089AGCACCCGGCCTCCAGGCTG  86 124 1047545 2862 2881 13151 13170GGGCACAGATCCCACCAGTC  65 125 1047561 2924 2943 13213 13232CCCTTCTTCGGCCTTAGAGG  62 126 1047577 3040 3059 13329 13348TTTTTCCTCAGCGACTAAAG  49 127 1047593 3060 3079 13349 13368GGGCGCAGCATTTGTCTTTA  59 128 1047609 N/A N/A  8778  8797GGCTTTTGAGATATCTTGTG  13 129 1047625 N/A N/A  9036  9055TGCCAGTTTAATGTACAGTT  72 130 1047641 N/A N/A  9072  9091ACCCAAGGACTCACCACCTT  89 131 1047657 N/A N/A  9212  9231ATGGAGCCTCAGGGATGAAA  69 132 1047673 N/A N/A  8382  8401CCCTGCTGTACTGACCTCGA  91 133 1047689 N/A N/A  8460  8479CCTGATCCTCAGTCCCAGTC  82 134 1047705 N/A N/A  8506  8525CGCTGCAGTGTCACGAAGGC  63 135 1047721 N/A N/A  8643  8662GGCAGATGTCAAGCTCTCAC  83 136 1047737 N/A N/A  3963  3982TCCTCACTTCTGCCTCACAG  66 137 1047753 N/A N/A  4095  4114AAGGATAGTGCCCCATCAAG  69 138 1047769 N/A N/A  4265  4284CAGTCACAAAGCCCAGCCAT  64 139 1047785 N/A N/A  4324  4343CCGCTTCCAACTCCTCCTTT  49 140 1047801 N/A N/A  4361  4380CCCAGAATCCAATCTCCCTC  85 141 1047817 N/A N/A  4419  4438AGGCAGTCACCTGTGCTTTG  96 142 1047833 N/A N/A  4526  4545GATTCCTCTGATCCCAGGTA  63 143 1047849 N/A N/A  4705  4724GCCTTAACTCATTACTAAGG  82 144 1047865 N/A N/A  4807  4826AGAGACCACCCCCACCCAGG  91 145 1047881 N/A N/A  4910  4929TGGTTTCATCCTGGAGCCTG  60 146 1047897 N/A N/A  5012  5031GGTGGGTGGCCATCAATCCT  71 147 1047913 N/A N/A  5168  5187CATCTGCTTCCTGGAGTGGC  31 148 1047929 N/A N/A  5400  5419TCTTTCATTTCCTGTCTCTA  81 149 1047945 N/A N/A  5561  5580CCCGAACCTCCTGACCAGGG 117 150 1047961 N/A N/A  5800  5819GCTTCCTCCACCCTCCTTCC  75 151 1047977 N/A N/A  5935  5954CAGCTACTACTAATAATAGC 104 152 1047993 N/A N/A  6040  6059TTAGTTAACCTCTCTGGACT  79 153 1048009 N/A N/A  6147  6166TAACCCAAAACAGACTGGCA 101 154 1048025 N/A N/A  6282  6301TCCCACACTACATATAAGCT  83 155 1048041 N/A N/A  6330  6349GCCTGTCCTGCCTAGCCCAA  80 156 1048057 N/A N/A  6464  6483GCCACTCACACTCCTCAGCT  79 157 1048073 N/A N/A  6548  6567TCTAATAGCCCTTTCTCCCC  88 158 1048089 N/A N/A  6961  6980AGAATCCAGAACCTTCCACA  88 159 1048105 N/A N/A  7074  7093CTGGGACTTTTCCCAACAAC  86 160 1048121 N/A N/A  7418  7437AGGCCCCGCCCTCGACCCAG  95 161 1048137 N/A N/A  7915  7934AAACATCTAGTGACTGCCTG  87 162 1048153 N/A N/A  8061  8080AAGGAGGCAGAAGAGATGGG  59 163 1048169 N/A N/A  8286  8305CTGGGACACCCCTAGGCTGG  68 164 1048185 N/A N/A  9374  9393TTAGCTCCCCCCTCCCCCCG  84 165 1048201 N/A N/A  9536  9555TGGCAGTATTACCTCTACTA  43 166 1048217 N/A N/A  9611  9630TCCTGTCCCCTTTCCTCTTT  63 167 1048233 N/A N/A  9792  9811GCCACCAACCAGCCACATGA  89 168 1048249 N/A N/A 10005 10024CTGTAATCCCCTTACTCGGG  94 169 1048265 N/A N/A 10636 10655CTGCTCTGTCTTCTGGCCTG  65 170 1048281 N/A N/A 10735 10754GAGTATGAGAACCTATGCAA  53 171 1048297 N/A N/A 10794 10813AGGGCATGAGCCATCCTCTC  78 172 1048313 N/A N/A 10944 10963CTGACTGGGCCCAAATCCCT  95 173 1048329 N/A N/A 11074 11093ACATTCAGTTTCCTTGCTCT  89 174 1048345 N/A N/A 11228 11247AGCTACTAACTTTAATTCTC  82 175 1048361 N/A N/A 11330 11349GCCAAATCCCCTCTTCCCAT  19 176 1048377 N/A N/A 11491 11510CCAGGTCTTACTTTTCTTGA  85 177

TABLE 4Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 GFAP SEQ Compound Start Stop Start Stop RNA ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047146   58   77  3506  3525CTCCTCTCCATCCTGCTCTG  94 178 1047162  138  157  3586  3605GACGGCGGCCAGGAGCCAGG 115 179 1047178  274  293  3722  3741AGCTCCATCATCTCTGCCCG 117 180 1047194  495  514  3943  3962TGGCCAGGTCCTGTGCCAGA  67* 181 1047210  588  607  5177  5196GGGTGGCTTCATCTGCTTCC  18* 182 1047226  656  675  5245  5264GATCTTCCTCAAGAACCGGA  62 183 1047242  822  841  5708  5727AGCGGTACCACTCTTCGGCT 113 184 1047258 1052 1071  7624  7643CCCCTCTTCCTCCAGCCGCG  16 185 1047274 1253 1272 10867 10886GCCTTCTGACACAGACTTGG 113 186 1047290 1309 1328 N/A N/AATGACCTCTCCATCCCGCAT 108 187 1047306 1353 1372 11642 11661TCCTGCCTCACATCACATCC  75 188 1047322 1499 1518 11788 11807GCAAGCTGACCTAGGGACAG  68 189 1047338 1632 1651 11921 11940TCCTCATTCTAACGCAAGCT  68 190 1047354 1666 1685 11955 11974GGTGCCCCCCGCCCTCCTCC  90 191 1047370 1740 1759 12029 12048CTCCGAGAGAACCTCCATCT  77 192 1047386 1793 1812 12082 12101TCTCAGTTTTCCTCCAGCAG  33 193 1047402 1888 1907 12177 12196AGCATAGGGATATCCCACCT  63 194 1047418 2063 2082 12352 12371GGCAGGTGACTGCCCCAGGT 106 195 1047434 2182 2201 12471 12490AAGTGCTGAGAATCAAGCTC 103 196 1047450 2239 2258 12528 12547GCCCCCCTCTATCCCTCCCA 103 197 1047466 2329 2348 12618 12637CTCCTCCATCTCTACCAGCG  52 198 1047482 2381 2400 12670 12689ACTGGTCACCCACAACCCCT  79 199 1047498 2461 2480 12750 12769CATCCCCTTTCTCTCCTGTT  74 200 1047514 2532 2551 12821 12840CGGCCTGGTATGACACAGCA  91 201 1047530 2782 2801 13071 13090GAGCACCCGGCCTCCAGGCT  80 202 1047546 2864 2883 13153 13172CTGGGCACAGATCCCACCAG 107 203 1047562 2927 2946 13216 13235GGACCCTTCTTCGGCCTTAG  69 204 1047578 3043 3062 13332 13351TTATTTTTCCTCAGCGACTA  75 205 1047594 3062 3081 13351 13370AAGGGCGCAGCATTTGTCTT  86 206 1047610 N/A N/A  8782  8801GTGAGGCTTTTGAGATATCT  33 207 1047626 N/A N/A  9038  9057TCTGCCAGTTTAATGTACAG 114 208 1047642 N/A N/A  9078  9097CTGCGCACCCAAGGACTCAC  96 209 1047658 N/A N/A  9257  9276CCTGAGGGAAGAATCCTCTG  86 210 1047674 N/A N/A  8390  8409CCACGAGGCCCTGCTGTACT 111 211 1047690 N/A N/A  8461  8480CCCTGATCCTCAGTCCCAGT  82 212 1047706 N/A N/A  8534  8553CCCTGGTATGATAGGCTCTG  62 213 1047722 N/A N/A  8645  8664AGGGCAGATGTCAAGCTCTC 110 214 1047738 N/A N/A  3968  3987TCCCCTCCTCACTTCTGCCT  91 215 1047754 N/A N/A  4097  4116GCAAGGATAGTGCCCCATCA  94 216 1047770 N/A N/A  4268  4287CCACAGTCACAAAGCCCAGC  84 217 1047786 N/A N/A  4325  4344TCCGCTTCCAACTCCTCCTT 107 218 1047802 N/A N/A  4362  4381CCCCAGAATCCAATCTCCCT  84 219 1047818 N/A N/A  4477  4496CCACCGCTTCACAGCTGTGC  77 220 1047834 N/A N/A  4528  4547GGGATTCCTCTGATCCCAGG  93 221 1047850 N/A N/A  4706  4725TGCCTTAACTCATTACTAAG  64 222 1047866 N/A N/A  4809  4828ACAGAGACCACCCCCACCCA  63 223 1047882 N/A N/A  4959  4978CTGACCTGTCTATAGGCAGC  89* 224 1047898 N/A N/A  5085  5104GCCTTACCCCTCCTTCTGGG  100 225 1047914 N/A N/A  5268  5287TGCCCTGGCCTCACCTCCTC  98 226 1047930 N/A N/A  5401  5420GTCTTTCATTTCCTGTCTCT  55 227 1047946 N/A N/A  5562  5581TCCCGAACCTCCTGACCAGG 117 228 1047962 N/A N/A  5803  5822CCAGCTTCCTCCACCCTCCT 137 229 1047978 N/A N/A  5936  5955TCAGCTACTACTAATAATAG  91 230 1047994 N/A N/A  6077  6096AACTCTACCACTTAGGAGCT 130 231 1048010 N/A N/A  6148  6167GTAACCCAAAACAGACTGGC  85 232 1048026 N/A N/A  6283  6302CTCCCACACTACATATAAGC  82 233 1048042 N/A N/A  6331  6350TGCCTGTCCTGCCTAGCCCA  72 234 1048058 N/A N/A  6467  6486TCTGCCACTCACACTCCTCA  95 235 1048074 N/A N/A  6549  6568TTCTAATAGCCCTTTCTCCC  83 236 1048090 N/A N/A  6990  7009TCAGCAAGCGAATGAATGAA 157 237 1048106 N/A N/A  7075  7094GCTGGGACTTTTCCCAACAA  91 238 1048122 N/A N/A  7449  7468AGGCCCCGCCTCTAGCCCGG 110 239 1048138 N/A N/A  7920  7939TCATCAAACATCTAGTGACT  91 240 1048154 N/A N/A  8112  8131TCTATCTGAAGGAAGATGGA  90 241 1048170 N/A N/A  8329  8348GTGATCCTGAAAGAAAGCAG  68 242 1048186 N/A N/A  9375  9394TTTAGCTCCCCCCTCCCCCC 126 243 1048202 N/A N/A  9566  9585TGCTTTAGTGACCTGTGACT  74 244 1048218 N/A N/A  9614  9633CTTTCCTGTCCCCTTTCCTC 134 245 1048234 N/A N/A  9793  9812AGCCACCAACCAGCCACATG  83 246 1048250 N/A N/A 10006 10025CCTGTAATCCCCTTACTCGG  92 247 1048266 N/A N/A 10652 10671GCTGCCAGAGTCCTGGCTGC  81 248 1048282 N/A N/A 10742 10761CCATCATGAGTATGAGAACC  88 249 1048298 N/A N/A 10814 10833GAGGCCTCTCATGGACTTTC  84 250 1048314 N/A N/A 10953 10972AGCCAGAGCCTGACTGGGCC  80 251 1048330 N/A N/A 11080 11099GGAATTACATTCAGTTTCCT  72 252 1048346 N/A N/A 11268 11287CTCCCCATCCCCAACTGTGT 107 253 1048362 N/A N/A 11331 11350CGCCAAATCCCCTCTTCCCA 106 254 1048378 N/A N/A 11492 11511CCCAGGTCTTACTTTTCTTG  80 255

TABLE 5Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 GFAP SEQ Compound Start Stop Start Stop RNA ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047147   61   80  3509  3528CGTCTCCTCTCCATCCTGCT  74 256 1047163  139  158  3587  3606AGACGGCGGCCAGGAGCCAG 100 257 1047179  275  294  3723  3742GAGCTCCATCATCTCTGCCC 116 258 1047195  500  519  3948  3967CACAGTGGCCAGGTCCTGTG  70* 259 1047211  589  608  5178  5197AGGGTGGCTTCATCTGCTTC  37* 260 1047227  657  676  5246  5265GGATCTTCCTCAAGAACCGG  90 261 1047243  823  842  5709  5728GAGCGGTACCACTCTTCGGC  83 262 1047259 1054 1073  7626  7645TGCCCCTCTTCCTCCAGCCG  63 263 1047275 1254 1273 10868 10887GGCCTTCTGACACAGACTTG 116 264 1047291 1311 1330 N/A N/ATAATGACCTCTCCATCCCGC  91 265 1047307 1354 1373 11643 11662GTCCTGCCTCACATCACATC  88 266 1047323 1524 1543 11813 11832CCTGATACTGACGGAGCCTA  56 267 1047339 1633 1652 11922 11941CTCCTCATTCTAACGCAAGC 116 268 1047355 1668 1687 11957 11976TAGGTGCCCCCCGCCCTCCT  68 269 1047371 1754 1773 12043 12062ACAGTTCCCAGATACTCCGA  80 270 1047387 1794 1813 12083 12102GTCTCAGTTTTCCTCCAGCA  12 271 1047403 1966 1985 12255 12274CCAATCTATAATCCCAGCTA  89 272 1047419 2065 2084 12354 12373TGGGCAGGTGACTGCCCCAG 110 273 1047435 2192 2211 12481 12500CAGATCCCCCAAGTGCTGAG  87 274 1047451 2240 2259 12529 12548AGCCCCCCTCTATCCCTCCC  97 275 1047467 2335 2354 12624 12643TGCCTCCTCCTCCATCTCTA  72 276 1047483 2384 2403 12673 12692GCAACTGGTCACCCACAACC  49 277 1047499 2462 2481 12751 12770ACATCCCCTTTCTCTCCTGT  77 278 1047515 2675 2694 12964 12983TTTGTGTGTGAGTAAGAAGG  49 279 1047531 2785 2804 13074 13093CCTGAGCACCCGGCCTCCAG  80 280 1047547 2865 2884 13154 13173TCTGGGCACAGATCCCACCA  85 281 1047563 2928 2947 13217 13236AGGACCCTTCTTCGGCCTTA  70 282 1047579 3044 3063 13333 13352TTTATTTTTCCTCAGCGACT  47 283 1047611 N/A N/A  8826  8845TTCCATTTACAATCTGGTGA  93 285 1047627 N/A N/A  9040  9059GCTCTGCCAGTTTAATGTAC 128 286 1047643 N/A N/A  9079  9098ACTGCGCACCCAAGGACTCA  77 287 1047659 N/A N/A  9283  9302ACTTTATTCACTGCAAGAGC  65 288 1047675 N/A N/A  8398  8417TGCCCTTCCCACGAGGCCCT  53 289 1047691 N/A N/A  8462  8481GCCCTGATCCTCAGTCCCAG  92 290 1047707 N/A N/A  8535  8554ACCCTGGTATGATAGGCTCT  46 291 1047723 N/A N/A  8662  8681CTCAGGGATCTGCAGACAGG  81 292 1047739 N/A N/A  3969  3988ATCCCCTCCTCACTTCTGCC  85* 293 1047755 N/A N/A  4119  4138GTCCCTCCCATCATGTTGGG  83 294 1047771 N/A N/A  4270  4289GCCCACAGTCACAAAGCCCA  68 295 1047787 N/A N/A  4327  4346TCTCCGCTTCCAACTCCTCC 109 296 1047803 N/A N/A  4363  4382ACCCCAGAATCCAATCTCCC  81 297 1047819 N/A N/A  4503  4522ACCTTTTGAAATGAATTTTA  65 298 1047835 N/A N/A  4588  4607CTCCTGCACTTGAAGGCACA 101 299 1047851 N/A N/A  4707  4726TTGCCTTAACTCATTACTAA  77 300 1047867 N/A N/A  4811  4830TCACAGAGACCACCCCCACC  97 301 1047883 N/A N/A  4964  4983CCTCCCTGACCTGTCTATAG 103* 302 1047899 N/A N/A  5086  5105TGCCTTACCCCTCCTTCTGG  76 303 1047915 N/A N/A  5270  5289TCTGCCCTGGCCTCACCTCC 102 304 1047931 N/A N/A  5403  5422TTGTCTTTCATTTCCTGTCT  67 305 1047947 N/A N/A  5563  5582TTCCCGAACCTCCTGACCAG 131 306 1047963 N/A N/A  5804  5823CCCAGCTTCCTCCACCCTCC  58 307 1047979 N/A N/A  5937  5956ATCAGCTACTACTAATAATA 128 308 1047995 N/A N/A  6078  6097CAACTCTACCACTTAGGAGC  97 309 1048011 N/A N/A  6150  6169CAGTAACCCAAAACAGACTG  85 310 1048027 N/A N/A  6284  6303GCTCCCACACTACATATAAG  48 311 1048043 N/A N/A  6352  6371CTTCTCTTCCTGTCCACAGC  93 312 1048059 N/A N/A  6471  6490GGCTTCTGCCACTCACACTC  85 313 1048075 N/A N/A  6550  6569GTTCTAATAGCCCTTTCTCC 124 314 1048091 N/A N/A  6991  7010GTCAGCAAGCGAATGAATGA  85 315 1048107 N/A N/A  7109  7128CAGCACCCCAGTTAACCCCA  73 316 1048123 N/A N/A  7450  7469CAGGCCCCGCCTCTAGCCCG  88 317 1048139 N/A N/A  7928  7947CCATTCAGTCATCAAACATC  76 318 1048155 N/A N/A  8120  8139GGCGCATGTCTATCTGAAGG  76 319 1048171 N/A N/A  8330  8349GGTGATCCTGAAAGAAAGCA 103 320 1048187 N/A N/A  9391  9410TGCAAGTAAAAAGTAATTTA  69 321 1048203 N/A N/A  9568  9587TGTGCTTTAGTGACCTGTGA  31 322 1048219 N/A N/A  9619  9638GGTCCCTTTCCTGTCCCCTT  53 323 1048235 N/A N/A  9794  9813TAGCCACCAACCAGCCACAT  97 324 1048251 N/A N/A 10007 10026ACCTGTAATCCCCTTACTCG  89 325 1048267 N/A N/A 10657 10676GTGCTGCTGCCAGAGTCCTG  35 326 1048283 N/A N/A 10750 10769CCCCCTCCCCATCATGAGTA 100 327 1048299 N/A N/A 10841 10860GGCTGGTTTCTGCAGATGTG  87 328 1048315 N/A N/A 10954 10973CAGCCAGAGCCTGACTGGGC 188 329 1048331 N/A N/A 11086 11105GGAAACGGAATTACATTCAG 130 330 1048347 N/A N/A 11269 11288CCTCCCCATCCCCAACTGTG  99 331 1048363 N/A N/A 11332 11351ACGCCAAATCCCCTCTTCCC  93 332 1048379 N/A N/A 11501 11520CCCCGACTTCCCAGGTCTTA  91 333

TABLE 6Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 GFAP SEQ Compound Start Stop Start Stop RNA ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047148  112  131  3560  3579ACCATCATCTCCCCTGAGGA 116 334 1047164  150  169  3598  3617TGCCAGGACCCAGACGGCGG  96 335 1047180  277  296  3725  3744TTGAGCTCCATCATCTCTGC 123 336 1047196  503  522  3951  3970CCTCACAGTGGCCAGGTCCT  27* 337 1047212  615  634  5204  5223TCTTCCTCTCCAGATCCAGA  77 338 1047228  658  677  5247  5266TGGATCTTCCTCAAGAACCG  75 339 1047244  824  843  5710  5729GGAGCGGTACCACTCTTCGG  74 340 1047260 1058 1077  7630  7649GCTCTGCCCCTCTTCCTCCA  82 341 1047276 1255 1274 10869 10888TGGCCTTCTGACACAGACTT 137 342 1047292 1312 1331 N/A N/ATTAATGACCTCTCCATCCCG  81 343 1047308 1356 1375 11645 11664GGGTCCTGCCTCACATCACA  69 344 1047324 1526 1545 11815 11834GGCCTGATACTGACGGAGCC  73 345 1047340 1634 1653 11923 11942CCTCCTCATTCTAACGCAAG  63 346 1047356 1672 1691 11961 11980GTAGTAGGTGCCCCCCGCCC  84 347 1047372 1755 1774 12044 12063CACAGTTCCCAGATACTCCG  49 348 1047388 1795 1814 12084 12103AGTCTCAGTTTTCCTCCAGC  24 349 1047404 1999 2018 12288 12307GGAGAACAACCCTCTGAGCT  75 350 1047420 2101 2120 12390 12409CACTTGAGTCATCGCTCAGG 107 351 1047436 2193 2212 12482 12501ACAGATCCCCCAAGTGCTGA  73 352 1047452 2241 2260 12530 12549CAGCCCCCCTCTATCCCTCC  88 353 1047468 2337 2356 12626 12645ATTGCCTCCTCCTCCATCTC  93 354 1047484 2395 2414 12684 12703AGAGGCCAAGTGCAACTGGT  80 355 1047500 2463 2482 12752 12771TACATCCCCTTTCTCTCCTG  47 356 1047516 2699 2718 12988 13007CACTACCTAGAATACTGGGT  92 357 1047532 2794 2813 13083 13102CGTGTCAGCCCTGAGCACCC 129 358 1047548 2866 2885 13155 13174CTCTGGGCACAGATCCCACC 141 359 1047564 2931 2950 13220 13239GGAAGGACCCTTCTTCGGCC  67 360 1047580 3045 3064 13334 13353CTTTATTTTTCCTCAGCGAC  35 361 1047612 N/A N/A  8827  8846GTTCCATTTACAATCTGGTG  60 363 1047628 N/A N/A  9053  9072TTACCACTAACAAGCTCTGC  76 364 1047644 N/A N/A  9081  9100CCACTGCGCACCCAAGGACT  89 365 1047660 N/A N/A  9290  9309ACATAAAACTTTATTCACTG  74 366 1047676 N/A N/A  8399  8418GTGCCCTTCCCACGAGGCCC 176 367 1047692 N/A N/A  8463  8482TGCCCTGATCCTCAGTCCCA  81 368 1047708 N/A N/A  8536  8555TACCCTGGTATGATAGGCTC  60 369 1047724 N/A N/A  8669  8688GTGCTTGCTCAGGGATCTGC  56 370 1047740 N/A N/A  3971  3990CCATCCCCTCCTCACTTCTG  85* 371 1047756 N/A N/A  4120  4139GGTCCCTCCCATCATGTTGG  76 372 1047772 N/A N/A  4272  4291CTGCCCACAGTCACAAAGCC 103 373 1047788 N/A N/A  4328  4347TTCTCCGCTTCCAACTCCTC 133 374 1047804 N/A N/A  4364  4383CACCCCAGAATCCAATCTCC  78 375 1047820 N/A N/A  4507  4526AACCACCTTTTGAAATGAAT 118 376 1047836 N/A N/A  4596  4615CACATGTCCTCCTGCACTTG 108 377 1047852 N/A N/A  4708  4727TTTGCCTTAACTCATTACTA  91 378 1047868 N/A N/A  4812  4831GTCACAGAGACCACCCCCAC 108 379 1047884 N/A N/A  4965  4984ACCTCCCTGACCTGTCTATA  83 380 1047900 N/A N/A  5087  5106TTGCCTTACCCCTCCTTCTG 102 381 1047916 N/A N/A  5282  5301AGCTTTCCTCCCTCTGCCCT  92 382 1047932 N/A N/A  5405  5424GTTTGTCTTTCATTTCCTGT  82 383 1047948 N/A N/A  5564  5583GTTCCCGAACCTCCTGACCA 103 384 1047964 N/A N/A  5805  5824TCCCAGCTTCCTCCACCCTC  78 385 1047980 N/A N/A  5938  5957TATCAGCTACTACTAATAAT 106 386 1047996 N/A N/A  6079  6098CCAACTCTACCACTTAGGAG  96 387 1048012 N/A N/A  6151  6170TCAGTAACCCAAAACAGACT 124 388 1048028 N/A N/A  6285  6304GGCTCCCACACTACATATAA 134 389 1048044 N/A N/A  6353  6372TCTTCTCTTCCTGTCCACAG  93 390 1048060 N/A N/A  6473  6492GTGGCTTCTGCCACTCACAC 129 391 1048076 N/A N/A  6556  6575CCCTGGGTTCTAATAGCCCT  89 392 1048092 N/A N/A  6993  7012TGGTCAGCAAGCGAATGAAT  81 393 1048108 N/A N/A  7111  7130AGCAGCACCCCAGTTAACCC 104 394 1048124 N/A N/A  7451  7470CCAGGCCCCGCCTCTAGCCC  89 395 1048140 N/A N/A  7932  7951CCATCCATTCAGTCATCAAA  70 396 1048156 N/A N/A  8142  8161GGCTTGAGTGTTATCTGGGA  74 397 1048172 N/A N/A  8332  8351ATGGTGATCCTGAAAGAAAG  87 398 1048188 N/A N/A  9392  9411ATGCAAGTAAAAAGTAATTT  61 399 1048204 N/A N/A  9581  9600TCTGCCATTTATCTGTGCTT  61 400 1048220 N/A N/A  9621  9640TAGGTCCCTTTCCTGTCCCC  60 401 1048236 N/A N/A  9795  9814TTAGCCACCAACCAGCCACA 106 402 1048252 N/A N/A 10009 10028GCACCTGTAATCCCCTTACT  76 403 1048268 N/A N/A 10658 10677AGTGCTGCTGCCAGAGTCCT  80 404 1048284 N/A N/A 10751 10770CCCCCCTCCCCATCATGAGT  82 405 1048300 N/A N/A 10921 10940TACCTCTCCATCCCGCATCT 109 406 1048316 N/A N/A 10976 10995TGGCCTTGAGAATCCCTGGG  99 407 1048332 N/A N/A 11090 11109CTGAGGAAACGGAATTACAT  83 408 1048348 N/A N/A 11272 11291AGCCCTCCCCATCCCCAACT 121 409 1048364 N/A N/A 11333 11352TACGCCAAATCCCCTCTTCC  92 410 1048380 N/A N/A 11504 11523AGTCCCCGACTTCCCAGGTC 116 411

TABLE 7Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 GFAP SEQ Compound Start Stop Start Stop RNA ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047149  113  132  3561  3580CACCATCATCTCCCCTGAGG 102 412 1047165  151  170  3599  3618GTGCCAGGACCCAGACGGCG 120 413 1047181  279  298  3727  3746CATTGAGCTCCATCATCTCT  92 414 1047197  525  544 N/A N/ATGGTTTCATCCTGGAGCTTC  39* 415 1047213  617  636  5206  5225AATCTTCCTCTCCAGATCCA  79 416 1047229  659  678  5248  5267GTGGATCTTCCTCAAGAACC  83 417 1047245  825  844  5711  5730TGGAGCGGTACCACTCTTCG  77 418 1047261 1060 1079  7632  7651AGGCTCTGCCCCTCTTCCTC  78 419 1047277 1257 1276 10871 10890GGTGGCCTTCTGACACAGAC  98 420 1047293 1313 1332 N/A N/ACTTAATGACCTCTCCATCCC 105 421 1047309 1357 1376 11646 11665TGGGTCCTGCCTCACATCAC 154 422 1047325 1529 1548 11818 11837GCAGGCCTGATACTGACGGA  46 423 1047341 1635 1654 11924 11943TCCTCCTCATTCTAACGCAA  53 424 1047357 1684 1703 11973 11992GTGGAGGGCGATGTAGTAGG  42 425 1047373 1756 1775 12045 12064GCACAGTTCCCAGATACTCC  33 426 1047389 1812 1831 12101 12120CTTCCCTTTCCTGTCTGAGT  64 427 1047405 2003 2022 12292 12311TCTAGGAGAACAACCCTCTG 160 428 1047421 2103 2122 12392 12411GACACTTGAGTCATCGCTCA  63 429 1047437 2194 2213 12483 12502AACAGATCCCCCAAGTGCTG  85 430 1047453 2242 2261 12531 12550GCAGCCCCCCTCTATCCCTC 173 431 1047469 2341 2360 12630 12649CCCAATTGCCTCCTCCTCCA  53 432 1047485 2407 2426 12696 12715TTCCCACAATCCAGAGGCCA 101 433 1047501 2464 2483 12753 12772ATACATCCCCTTTCTCTCCT  63 434 1047517 2701 2720 12990 13009GGCACTACCTAGAATACTGG  73 435 1047533 2833 2852 13122 13141GTCTGCTCAGTCAAAGCAGA  93 436 1047549 2877 2896 13166 13185CCCAGTCCCATCTCTGGGCA 134 437 1047565 2932 2951 13221 13240GGGAAGGACCCTTCTTCGGC  66 438 1047581 3046 3065 13335 13354TCTTTATTTTTCCTCAGCGA  41 439 1047613 N/A N/A  8828  8847CGTTCCATTTACAATCTGGT  47 441 1047629 N/A N/A  9056  9075CCTTTACCACTAACAAGCTC 161 442 1047645 N/A N/A  9086  9105CAGCTCCACTGCGCACCCAA  85 443 1047661 N/A N/A  9300  9319AGAGCAGGGAACATAAAACT  69 444 1047677 N/A N/A  8400  8419AGTGCCCTTCCCACGAGGCC  85 445 1047693 N/A N/A  8464  8483TTGCCCTGATCCTCAGTCCC 117 446 1047709 N/A N/A  8547  8566CCACCTAGAAGTACCCTGGT  75 447 1047725 N/A N/A  8688  8707GAAAACACTCAGAAGGGCAG 101 448 1047741 N/A N/A  3972  3991CCCATCCCCTCCTCACTTCT 133 449 1047757 N/A N/A  4122  4141CTGGTCCCTCCCATCATGTT 136 450 1047773 N/A N/A  4273  4292GCTGCCCACAGTCACAAAGC 126 451 1047789 N/A N/A  4334  4353TCAACCTTCTCCGCTTCCAA  66 452 1047805 N/A N/A  4369  4388TTCTTCACCCCAGAATCCAA 123 453 1047821 N/A N/A  4508  4527TAACCACCTTTTGAAATGAA  75 454 1047837 N/A N/A  4657  4676CTGCTCACACAGGCGCATCC  97 455 1047853 N/A N/A  4709  4728TTTTGCCTTAACTCATTACT 142 456 1047869 N/A N/A  4813  4832TGTCACAGAGACCACCCCCA  87 457 1047885 N/A N/A  4968  4987TCCACCTCCCTGACCTGTCT  84 458 1047901 N/A N/A  5090  5109GCCTTGCCTTACCCCTCCTT  86 459 1047917 N/A N/A  5284  5303TGAGCTTTCCTCCCTCTGCC  93 460 1047933 N/A N/A  5417  5436TAGTGTCTTTCTGTTTGTCT  82 461 1047949 N/A N/A  5565  5584AGTTCCCGAACCTCCTGACC  74 462 1047965 N/A N/A  5806  5825CTCCCAGCTTCCTCCACCCT 101 463 1047981 N/A N/A  5939  5958GTATCAGCTACTACTAATAA 137 464 1047997 N/A N/A  6080  6099TCCAACTCTACCACTTAGGA 112 465 1048013 N/A N/A  6152  6171CTCAGTAACCCAAAACAGAC  79 466 1048029 N/A N/A  6287  6306CTGGCTCCCACACTACATAT  86 467 1048045 N/A N/A  6354  6373TTCTTCTCTTCCTGTCCACA  92 468 1048061 N/A N/A  6474  6493AGTGGCTTCTGCCACTCACA  88 469 1048077 N/A N/A  6557  6576ACCCTGGGTTCTAATAGCCC 104 470 1048093 N/A N/A  7008  7027ACCTAGCACAACACCTGGTC  77 471 1048109 N/A N/A  7318  7337GCGCTCACCGTGCCGCGCAG  85 472 1048125 N/A N/A  7497  7516GAGCCCCGACCCGACTTGGG  55 473 1048141 N/A N/A  7943  7962GTTGAATCCATCCATCCATT  73 474 1048157 N/A N/A  8195  8214AGCTTTTTCCCCAGCAGCCA  91 475 1048173 N/A N/A  8333  8352AATGGTGATCCTGAAAGAAA  80 476 1048189 N/A N/A  9415  9434AGCTAAGAATCATTTCAGGG  77 477 1048205 N/A N/A  9584  9603CTCTCTGCCATTTATCTGTG  67 478 1048221 N/A N/A  9622  9641ATAGGTCCCTTTCCTGTCCC  67 479 1048237 N/A N/A  9796  9815CTTAGCCACCAACCAGCCAC 123 480 1048253 N/A N/A 10010 10029CGCACCTGTAATCCCCTTAC  97 481 1048269 N/A N/A 10666 10685ATCCCAATAGTGCTGCTGCC  79 482 1048285 N/A N/A 10755 10774CGCACCCCCCTCCCCATCAT 109 483 1048301 N/A N/A 10925 10944TCCTTACCTCTCCATCCCGC 106 484 1048317 N/A N/A 10993 11012GGCTTTCCTCCATGGCCTGG  93 485 1048333 N/A N/A 11092 11111GACTGAGGAAACGGAATTAC  86 486 1048349 N/A N/A 11278 11297ATGGAAAGCCCTCCCCATCC  86 487 1048365 N/A N/A 11334 11353ATACGCCAAATCCCCTCTTC 105 488 1048381 N/A N/A 11505 11524AAGTCCCCGACTTCCCAGGT  79 489

TABLE 8Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047150  115  134  3563  3582CCCACCATCATCTCCCCTGA  93 490 1047166  156  175  3604  3623GGCGGGTGCCAGGACCCAGA  79 491 1047182  358  377  3806  3825AGCTGGTTCAGCTCAGCAGC 114 492 1047198  528  547  4913  4932GGTTGGTTTCATCCTGGAGC  19* 493 1047214  619  638  5208  5227TCAATCTTCCTCTCCAGATC 115 494 1047230  680  699 N/A N/AGAGTTCCCGAACCTCCTCCT  85 495 1047246  838  857 N/A N/AAGGTCTGCAAACTTGGAGCG  90 496 1047262 1064 1083  7636  7655CTTGAGGCTCTGCCCCTCTT  88 497 1047278 1258 1277 10872 10891AGGTGGCCTTCTGACACAGA  94 498 1047294 1314 1333 N/A N/ACCTTAATGACCTCTCCATCC 154 499 1047310 1358 1377 11647 11666GTGGGTCCTGCCTCACATCA  88 500 1047326 1530 1549 11819 11838GGCAGGCCTGATACTGACGG 103 501 1047342 1636 1655 11925 11944TTCCTCCTCATTCTAACGCA  66 502 1047358 1685 1704 11974 11993TGTGGAGGGCGATGTAGTAG  50 503 1047374 1757 1776 12046 12065GGCACAGTTCCCAGATACTC  40 504 1047390 1813 1832 12102 12121CCTTCCCTTTCCTGTCTGAG  81 505 1047406 2004 2023 12293 12312GTCTAGGAGAACAACCCTCT  96 506 1047422 2112 2131 12401 12420GTGGACTGAGACACTTGAGT  54 507 1047438 2201 2220 12490 12509CGTACACAACAGATCCCCCA  61 508 1047454 2243 2262 12532 12551GGCAGCCCCCCTCTATCCCT  57 509 1047470 2342 2361 12631 12650TCCCAATTGCCTCCTCCTCC  66 510 1047486 2413 2432 12702 12721CCTTAATTCCCACAATCCAG 147 511 1047502 2465 2484 12754 12773GATACATCCCCTTTCTCTCC  67 512 1047518 2702 2721 12991 13010GGGCACTACCTAGAATACTG  43 513 1047534 2843 2862 13132 13151CTGCTCACCAGTCTGCTCAG  97 514 1047550 2878 2897 13167 13186TCCCAGTCCCATCTCTGGGC 106 515 1047566 2933 2952 13222 13241AGGGAAGGACCCTTCTTCGG  71 516 1047582 3047 3066 13336 13355GTCTTTATTTTTCCTCAGCG   9 517 1047614 N/A N/A  8850  8869GCAGCTAACCGCGAGCCGGC 132 519 1047630 N/A N/A  9057  9076ACCTTTACCACTAACAAGCT  89 520 1047646 N/A N/A  9088  9107AGCAGCTCCACTGCGCACCC 116 521 1047662 N/A N/A  9311  9330ATTTAACATTAAGAGCAGGG  45 522 1047678 N/A N/A  8401  8420CAGTGCCCTTCCCACGAGGC 110 523 1047694 N/A N/A  8467  8486CCTTTGCCCTGATCCTCAGT  77 524 1047710 N/A N/A  8549  8568CCCCACCTAGAAGTACCCTG  99 525 1047726 N/A N/A  8689  8708AGAAAACACTCAGAAGGGCA 169 526 1047742 N/A N/A  3973  3992CCCCATCCCCTCCTCACTTC 100 527 1047758 N/A N/A  4124  4143TTCTGGTCCCTCCCATCATG  77 528 1047774 N/A N/A  4277  4296GCTCGCTGCCCACAGTCACA 123 529 1047790 N/A N/A  4338  4357GACATCAACCTTCTCCGCTT  67 530 1047806 N/A N/A  4375  4394CTCACTTTCTTCACCCCAGA 156 531 1047822 N/A N/A  4510  4529GGTAACCACCTTTTGAAATG  91 532 1047838 N/A N/A  4659  4678TTCTGCTCACACAGGCGCAT 148 533 1047854 N/A N/A  4712  4731GGCTTTTGCCTTAACTCATT  72 534 1047870 N/A N/A  4815  4834GCTGTCACAGAGACCACCCC  93 535 1047886 N/A N/A  4971  4990CCCTCCACCTCCCTGACCTG  59 536 1047902 N/A N/A  5092  5111CAGCCTTGCCTTACCCCTCC  70 537 1047918 N/A N/A  5285  5304TTGAGCTTTCCTCCCTCTGC  80 538 1047934 N/A N/A  5429  5448TTCCGTCTCCCTTAGTGTCT 105 539 1047950 N/A N/A  5566  5585GAGTTCCCGAACCTCCTGAC  70 540 1047966 N/A N/A  5813  5832GGATATTCTCCCAGCTTCCT  90 541 1047982 N/A N/A  5945  5964AGAACAGTATCAGCTACTAC 106 542 1047998 N/A N/A  6085  6104GGAAATCCAACTCTACCACT 106 543 1048014 N/A N/A  6153  6172GCTCAGTAACCCAAAACAGA  96 544 1048030 N/A N/A  6288  6307CCTGGCTCCCACACTACATA 116 545 1048046 N/A N/A  6356  6375CCTTCTTCTCTTCCTGTCCA 133 546 1048062 N/A N/A  6486  6505TGCTCAGACACCAGTGGCTT 109 547 1048078 N/A N/A  6567  6586GCCTGGCCTCACCCTGGGTT  88 548 1048094 N/A N/A  7023  7042CTGCCAGACCTCAGCACCTA  81 549 1048110 N/A N/A  7322  7341GGCCGCGCTCACCGTGCCGC 109 550 1048126 N/A N/A  7498  7517GGAGCCCCGACCCGACTTGG 124 551 1048142 N/A N/A  7944  7963GGTTGAATCCATCCATCCAT 105 552 1048158 N/A N/A  8196  8215TAGCTTTTTCCCCAGCAGCC  89 553 1048174 N/A N/A  8334  8353GAATGGTGATCCTGAAAGAA  81 554 1048190 N/A N/A  9462  9481AGTGGTCCTAAATATTCTAG  66 555 1048206 N/A N/A  9585  9604TCTCTCTGCCATTTATCTGT  91 556 1048222 N/A N/A  9623  9642CATAGGTCCCTTTCCTGTCC  71 557 1048238 N/A N/A  9799  9818CAACTTAGCCACCAACCAGC 120 558 1048254 N/A N/A 10576 10595GGCTTTCTGAAAACCCAGCA  72 559 1048270 N/A N/A 10673 10692CCCCCAAATCCCAATAGTGC 117 560 1048286 N/A N/A 10756 10775TCGCACCCCCCTCCCCATCA 106 561 1048302 N/A N/A 10926 10945CTCCTTACCTCTCCATCCCG 120 562 1048318 N/A N/A 10994 11013AGGCTTTCCTCCATGGCCTG  91 563 1048334 N/A N/A 11110 11129TAGAACAGCCTATGGAGGGA  52 564 1048350 N/A N/A 11300 11319GTTTCCTTTTACCAAGCTGG  34 565 1048366 N/A N/A 11335 11354GATACGCCAAATCCCCTCTT  94 566 1048382 N/A N/A 11508 11527GGGAAGTCCCCGACTTCCCA  92 567

TABLE 9Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047151  116  135  3564  3583CCCCACCATCATCTCCCCTG  89 568 1047167  173  192  3621  3640CATTCGAGCCAGGGAGAGGC  82 569 1047183  372  391  3820  3839GCTCCTTGGCCCGCAGCTGG 147 570 1047199  529  548  4914  4933AGGTTGGTTTCATCCTGGAG  47* 571 1047215  620  639  5209  5228CTCAATCTTCCTCTCCAGAT  93 572 1047231  683  702  5569  5588CTGGAGTTCCCGAACCTCCT 113 573 1047247  943  962  7300  7319AGAGACTCCAGGTCGCAGGT 104 574 1047263 1098 1117  7670  7689CCTGGTACTCCTGCAAGTGG 139 575 1047279 1260 1279 10874 10893TGAGGTGGCCTTCTGACACA 106 576 1047295 1319 1338 N/A N/AGGACTCCTTAATGACCTCTC  77 577 1047311 1370 1389 11659 11678AGAGGCCACCAGGTGGGTCC  88 578 1047327 1531 1550 11820 11839TGGCAGGCCTGATACTGACG  71 579 1047343 1637 1656 11926 11945CTTCCTCCTCATTCTAACGC  68 580 1047359 1714 1733 12003 12022ACAGTTTCCATAACAACAGG 137 581 1047375 1760 1779 12049 12068AAAGGCACAGTTCCCAGATA 106 582 1047391 1814 1833 12103 12122GCCTTCCCTTTCCTGTCTGA  34 583 1047407 2005 2024 12294 12313AGTCTAGGAGAACAACCCTC 113 584 1047423 2138 2157 12427 12446GATGGCATCCCTGGATGGCA 113 585 1047439 2219 2238 12508 12527GCACCTCATCCCTCTCCACG  75 586 1047455 2244 2263 12533 12552AGGCAGCCCCCCTCTATCCC  77 587 1047471 2343 2362 12632 12651ATCCCAATTGCCTCCTCCTC  96 588 1047487 2414 2433 12703 12722TCCTTAATTCCCACAATCCA 105 589 1047503 2466 2485 12755 12774GGATACATCCCCTTTCTCTC  48 590 1047519 2727 2746 13016 13035GCCTCAGTTTTACAATTGTA  90 591 1047535 2844 2863 13133 13152TCTGCTCACCAGTCTGCTCA  92 592 1047551 2880 2899 13169 13188CCTCCCAGTCCCATCTCTGG  90 593 1047567 2937 2956 13226 13245GGAGAGGGAAGGACCCTTCT 105 594 1047583 3048 3067 13337 13356TGTCTTTATTTTTCCTCAGC  10 595 1047615 N/A N/A  8851  8870GGCAGCTAACCGCGAGCCGG  92 597 1047631 N/A N/A  9058  9077CACCTTTACCACTAACAAGC 105 598 1047647 N/A N/A  9089  9108GAGCAGCTCCACTGCGCACC  87 599 1047663 N/A N/A  9312  9331TATTTAACATTAAGAGCAGG 106 600 1047679 N/A N/A  8402  8421CCAGTGCCCTTCCCACGAGG  78 601 1047695 N/A N/A  8469  8488TCCCTTTGCCCTGATCCTCA  74 602 1047711 N/A N/A  8551  8570AGCCCCACCTAGAAGTACCC  86 603 1047727 N/A N/A  8690  8709CAGAAAACACTCAGAAGGGC  93 604 1047743 N/A N/A  3974  3993TCCCCATCCCCTCCTCACTT  92 605 1047759 N/A N/A  4126  4145GTTTCTGGTCCCTCCCATCA  98 606 1047775 N/A N/A  4278  4297AGCTCGCTGCCCACAGTCAC 170 607 1047791 N/A N/A  4339  4358GGACATCAACCTTCTCCGCT  98 608 1047807 N/A N/A  4376  4395CCTCACTTTCTTCACCCCAG 104 609 1047823 N/A N/A  4512  4531CAGGTAACCACCTTTTGAAA  82 610 1047839 N/A N/A  4661  4680GCTTCTGCTCACACAGGCGC  96 611 1047855 N/A N/A  4713  4732GGGCTTTTGCCTTAACTCAT  89 612 1047871 N/A N/A  4833  4852TCAGTCTCCCTTGAGGCAGC  87 613 1047887 N/A N/A  4972  4991CCCCTCCACCTCCCTGACCT 109 614 1047903 N/A N/A  5093  5112TCAGCCTTGCCTTACCCCTC  96 615 1047919 N/A N/A  5286  5305GTTGAGCTTTCCTCCCTCTG  82 616 1047935 N/A N/A  5434  5453TCTCTTTCCGTCTCCCTTAG  83 617 1047951 N/A N/A  5724  5743GGCAGGGCTACCTTGGAGCG  98 618 1047967 N/A N/A  5814  5833AGGATATTCTCCCAGCTTCC  74 619 1047983 N/A N/A  5946  5965CAGAACAGTATCAGCTACTA  76 620 1047999 N/A N/A  6086  6105TGGAAATCCAACTCTACCAC 122 621 1048015 N/A N/A  6154  6173GGCTCAGTAACCCAAAACAG  82 622 1048031 N/A N/A  6290  6309TTCCTGGCTCCCACACTACA 106 623 1048047 N/A N/A  6362  6381GCCCTCCCTTCTTCTCTTCC  84 624 1048063 N/A N/A  6487  6506CTGCTCAGACACCAGTGGCT 111 625 1048079 N/A N/A  6569  6588TCGCCTGGCCTCACCCTGGG 101 626 1048095 N/A N/A  7024  7043GCTGCCAGACCTCAGCACCT 192 627 1048111 N/A N/A  7328  7347TGCCCTGGCCGCGCTCACCG  82 628 1048127 N/A N/A  7502  7521CCGCGGAGCCCCGACCCGAC  90 629 1048143 N/A N/A  7945  7964TGGTTGAATCCATCCATCCA 117 630 1048159 N/A N/A  8197  8216CTAGCTTTTTCCCCAGCAGC 100 631 1048175 N/A N/A  9316  9335CTAATATTTAACATTAAGAG 128 632 1048191 N/A N/A  9463  9482TAGTGGTCCTAAATATTCTA 106 633 1048207 N/A N/A  9586  9605TTCTCTCTGCCATTTATCTG  70 634 1048223 N/A N/A  9625  9644CACATAGGTCCCTTTCCTGT  74 635 1048239 N/A N/A  9800  9819CCAACTTAGCCACCAACCAG  96 636 1048255 N/A N/A 10577 10596TGGCTTTCTGAAAACCCAGC  83 637 1048271 N/A N/A 10674 10693GCCCCCAAATCCCAATAGTG 116 638 1048287 N/A N/A 10757 10776ATCGCACCCCCCTCCCCATC 142 639 1048303 N/A N/A 10928 10947CCCTCCTTACCTCTCCATCC 117 640 1048319 N/A N/A 10996 11015CCAGGCTTTCCTCCATGGCC  88 641 1048335 N/A N/A 11111 11130TTAGAACAGCCTATGGAGGG 115 642 1048351 N/A N/A 11301 11320AGTTTCCTTTTACCAAGCTG  54 643 1048367 N/A N/A 11337 11356CGGATACGCCAAATCCCCTC 101 644 1048383 N/A N/A 11539 11558CAGGTCCACCACCACGAGGC 138 645

TABLE 10Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047152  117  136  3565  3584CCCCCACCATCATCTCCCCT 104 646 1047168  175  194  3623  3642GGCATTCGAGCCAGGGAGAG  90 647 1047184  435  454  3883  3902GTTGATCGAGCCGCAGCCGC 104 648 1047200  530  549  4915  4934CAGGTTGGTTTCATCCTGGA  56* 649 1047216  621  640  5210  5229ACTCAATCTTCCTCTCCAGA  95 650 1047232  684  703  5570  5589CCTGGAGTTCCCGAACCTCC 126 651 1047248  946  965  7303  7322CGCAGAGACTCCAGGTCGCA 104 652 1047264 1100 1119  7672  7691GTCCTGGTACTCCTGCAAGT  89 653 1047280 1273 1292 10887 10906ACGATGTTCCTCTTGAGGTG  96 654 1047296 1320 1339 N/A N/ATGGACTCCTTAATGACCTCT 101 655 1047312 1372 1391 11661 11680GCAGAGGCCACCAGGTGGGT  64 656 1047328 1580 1599 11869 11888GGTGAGTTTCTTGTTAGTTG  29 657 1047344 1638 1657 11927 11946CCTTCCTCCTCATTCTAACG  73 658 1047360 1715 1734 12004 12023AACAGTTTCCATAACAACAG 115 659 1047376 1761 1780 12050 12069CAAAGGCACAGTTCCCAGAT  94 660 1047392 1815 1834 12104 12123GGCCTTCCCTTTCCTGTCTG 104 661 1047408 2021 2040 12310 12329TAGACTGATCAGGGTCAGTC 125 662 1047424 2148 2167 12437 12456CGTGCCCACAGATGGCATCC  87 663 1047440 2222 2241 12511 12530CCAGCACCTCATCCCTCTCC 111 664 1047456 2246 2265 12535 12554CCAGGCAGCCCCCCTCTATC  87 665 1047472 2345 2364 12634 12653CCATCCCAATTGCCTCCTCC  76 666 1047488 2417 2436 12706 12725ACTTCCTTAATTCCCACAAT 103 667 1047504 2468 2487 12757 12776ATGGATACATCCCCTTTCTC  73 668 1047520 2728 2747 13017 13036TGCCTCAGTTTTACAATTGT 109 669 1047536 2845 2864 13134 13153GTCTGCTCACCAGTCTGCTC 110 670 1047552 2883 2902 13172 13191GGCCCTCCCAGTCCCATCTC  85 671 1047568 2956 2975 13245 13264AAAGGACACCAAGTCTTGGG  69 672 1047584 3049 3068 13338 13357TTGTCTTTATTTTTCCTCAG  11 673 1047616 N/A N/A  8852  8871AGGCAGCTAACCGCGAGCCG  94 675 1047632 N/A N/A  9059  9078CCACCTTTACCACTAACAAG  89 676 1047648 N/A N/A  9101  9120TCAGAGGCCCCAGAGCAGCT  86 677 1047664 N/A N/A  9314  9333AATATTTAACATTAAGAGCA 105 678 1047680 N/A N/A  8404  8423CTCCAGTGCCCTTCCCACGA  86 679 1047696 N/A N/A  8471  8490GATCCCTTTGCCCTGATCCT  76 680 1047712 N/A N/A  8554  8573GCAAGCCCCACCTAGAAGTA  76 681 1047728 N/A N/A  8691  8710ACAGAAAACACTCAGAAGGG  92 682 1047744 N/A N/A  3984  4003AGGCCCCCCTTCCCCATCCC  79 683 1047760 N/A N/A  4138  4157GGCCCTGGGCCTGTTTCTGG  86 684 1047776 N/A N/A  4279  4298GAGCTCGCTGCCCACAGTCA 119 685 1047792 N/A N/A  4340  4359TGGACATCAACCTTCTCCGC 102 686 1047808 N/A N/A  4377  4396CCCTCACTTTCTTCACCCCA  71 687 1047824 N/A N/A  4513  4532CCAGGTAACCACCTTTTGAA 104 688 1047840 N/A N/A  4686  4705GTGCCTTATCAGGGTTGGTG  64 689 1047856 N/A N/A  4714  4733TGGGCTTTTGCCTTAACTCA  79 690 1047872 N/A N/A  4835  4854CCTCAGTCTCCCTTGAGGCA 104 691 1047888 N/A N/A  4976  4995CCCTCCCCTCCACCTCCCTG  79 692 1047904 N/A N/A  5094  5113CTCAGCCTTGCCTTACCCCT 121 693 1047920 N/A N/A  5313  5332TCTCCCTCTCTCAGTTGCAA  80 694 1047936 N/A N/A  5436  5455TGTCTCTTTCCGTCTCCCTT  76 695 1047952 N/A N/A  5738  5757CAGGCTGGCCCACAGGCAGG  96 696 1047968 N/A N/A  5815  5834GAGGATATTCTCCCAGCTTC  83 697 1047984 N/A N/A  5993  6012CACCTACTTCATAGTAAGGT 144 698 1048000 N/A N/A  6088  6107GTTGGAAATCCAACTCTACC  75 699 1048016 N/A N/A  6155  6174AGGCTCAGTAACCCAAAACA  97 700 1048032 N/A N/A  6298  6317CAGTGTCTTTCCTGGCTCCC  72 701 1048048 N/A N/A  6363  6382GGCCCTCCCTTCTTCTCTTC 106 702 1048064 N/A N/A  6491  6510CACCCTGCTCAGACACCAGT 112 703 1048080 N/A N/A  6570  6589CTCGCCTGGCCTCACCCTGG  80 704 1048096 N/A N/A  7028  7047GCGGGCTGCCAGACCTCAGC  82 705 1048112 N/A N/A  7335  7354CCCGTCCTGCCCTGGCCGCG  92 706 1048128 N/A N/A  7833  7852AAAAAGACTCAGTCCCTGAA 103 707 1048144 N/A N/A  7946  7965TTGGTTGAATCCATCCATCC  68 708 1048160 N/A N/A  8198  8217CCTAGCTTTTTCCCCAGCAG  71 709 1048176 N/A N/A  9317  9336ACTAATATTTAACATTAAGA  69 710 1048192 N/A N/A  9465  9484TCTAGTGGTCCTAAATATTC 108 711 1048208 N/A N/A  9593  9612TTCCTACTTCTCTCTGCCAT  80 712 1048224 N/A N/A  9633  9652GCTCAATACACATAGGTCCC  82 713 1048240 N/A N/A  9801  9820CCCAACTTAGCCACCAACCA 120 714 1048256 N/A N/A 10578 10597CTGGCTTTCTGAAAACCCAG 119 715 1048272 N/A N/A 10675 10694AGCCCCCAAATCCCAATAGT 116 716 1048288 N/A N/A 10759 10778CCATCGCACCCCCCTCCCCA  88 717 1048304 N/A N/A 10929 10948TCCCTCCTTACCTCTCCATC  73 718 1048320 N/A N/A 10997 11016CCCAGGCTTTCCTCCATGGC 113 719 1048336 N/A N/A 11112 11131CTTAGAACAGCCTATGGAGG  74 720 1048352 N/A N/A 11302 11321TAGTTTCCTTTTACCAAGCT  97 721 1048368 N/A N/A 11338 11357GCGGATACGCCAAATCCCCT  74 722 1048384 N/A N/A 11541 11560CCCAGGTCCACCACCACGAG 105 723

TABLE 11Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047153  119  138  3567  3586GCCCCCCACCATCATCTCCC  88 724 1047169  210  229  3658  3677CCAGGGAGAAATCCACCCGG  81 725 1047185  440  459  3888  3907GGTGAGTTGATCGAGCCGCA  70 726 1047201  539  558  4924  4943TTCCAGCCTCAGGTTGGTTT  29* 727 1047217  627  646  5216  5235CCAGCGACTCAATCTTCCTC  99 728 1047233  703  722  5589  5608TGCTGTCGGGCCAGCTGCTC 118 729 1047249  990 1009  7562  7581CCTCCTGCTCGCGCATCTGC  93 730 1047265 1101 1120  7673  7692GGTCCTGGTACTCCTGCAAG 129 731 1047281 1274 1293 10888 10907CACGATGTTCCTCTTGAGGT 104 732 1047297 1321 1340 N/A N/ATTGGACTCCTTAATGACCTC 110 733 1047313 1393 1412 11682 11701TCGGGCCCCTCATGAGACGG  99 734 1047329 1605 1624 11894 11913ATGCCCCTCCAGACTGCCCC 109 735 1047345 1640 1659 11929 11948CTCCTTCCTCCTCATTCTAA  89 736 1047361 1716 1735 12005 12024CAACAGTTTCCATAACAACA  62 737 1047377 1770 1789 12059 12078GAGGAAACTCAAAGGCACAG  82 738 1047393 1816 1835 12105 12124GGGCCTTCCCTTTCCTGTCT  64 739 1047409 2024 2043 12313 12332TCTTAGACTGATCAGGGTCA 124 740 1047425 2166 2185 12455 12474GCTCCCACCTGCCCACAGCG  79 741 1047441 2224 2243 12513 12532TCCCAGCACCTCATCCCTCT 109 742 1047457 2247 2266 12536 12555GCCAGGCAGCCCCCCTCTAT 100 743 1047473 2369 2388 12658 12677CAACCCCTACTTGTATGCCT  73 744 1047489 2429 2448 12718 12737AGAGGATGAGTCACTTCCTT 116 745 1047505 2469 2488 12758 12777CATGGATACATCCCCTTTCT  83 746 1047521 2748 2767 13037 13056CAGTGTCTTCACTTTGCTCG  69 747 1047537 2846 2865 13135 13154AGTCTGCTCACCAGTCTGCT 103 748 1047553 2884 2903 13173 13192GGGCCCTCCCAGTCCCATCT 105 749 1047569 2957 2976 13246 13265GAAAGGACACCAAGTCTTGG 117 750 1047585 3050 3069 13339 13358TTTGTCTTTATTTTTCCTCA  20 751 1047601 N/A N/A  8751  8770GATTTTCCCCGTCTTTGGTG  38 752 1047617 N/A N/A  8901  8920GTGAGGCTCACTCCCTGTCA  95 753 1047633 N/A N/A  9060  9079ACCACCTTTACCACTAACAA 108 754 1047649 N/A N/A  9107  9126GCTTGCTCAGAGGCCCCAGA  59 755 1047665 N/A N/A  9315  9334TAATATTTAACATTAAGAGC 135 756 1047681 N/A N/A  8406  8425GACTCCAGTGCCCTTCCCAC 109 757 1047697 N/A N/A  8473  8492TGGATCCCTTTGCCCTGATC  90 758 1047713 N/A N/A  8557  8576GCTGCAAGCCCCACCTAGAA  69 759 1047729 N/A N/A  8692  8711AACAGAAAACACTCAGAAGG  97 760 1047745 N/A N/A  3985  4004AAGGCCCCCCTTCCCCATCC 109 761 1047761 N/A N/A  4158  4177TGCGGGCATCAGATCCCCGG 138 762 1047777 N/A N/A  4313  4332TCCTCCTTTATATGGACACA 111 763 1047793 N/A N/A  4341  4360ATGGACATCAACCTTCTCCG  74 764 1047809 N/A N/A  4378  4397TCCCTCACTTTCTTCACCCC 100 765 1047825 N/A N/A  4514  4533CCCAGGTAACCACCTTTTGA  92 766 1047841 N/A N/A  4689  4708AAGGTGCCTTATCAGGGTTG  74 767 1047857 N/A N/A  4716  4735TGTGGGCTTTTGCCTTAACT 103 768 1047873 N/A N/A  4841  4860TACCTGCCTCAGTCTCCCTT 130 769 1047889 N/A N/A  4998  5017AATCCTTTCCTCCCTCCCCT 129 770 1047905 N/A N/A  5103  5122TCCCCATTCCTCAGCCTTGC 112 771 1047921 N/A N/A  5317  5336TGTCTCTCCCTCTCTCAGTT  88 772 1047937 N/A N/A  5438  5457CTTGTCTCTTTCCGTCTCCC 105 773 1047953 N/A N/A  5739  5758GCAGGCTGGCCCACAGGCAG  83 774 1047969 N/A N/A  5816  5835AGAGGATATTCTCCCAGCTT  80 775 1047985 N/A N/A  5994  6013GCACCTACTTCATAGTAAGG  65 776 1048001 N/A N/A  6089  6108AGTTGGAAATCCAACTCTAC  88 777 1048017 N/A N/A  6157  6176AGAGGCTCAGTAACCCAAAA  90 778 1048033 N/A N/A  6306  6325CCCCTCTACAGTGTCTTTCC 101 779 1048049 N/A N/A  6364  6383TGGCCCTCCCTTCTTCTCTT  89 780 1048065 N/A N/A  6497  6516GGCCCTCACCCTGCTCAGAC 143 781 1048081 N/A N/A  6571  6590CCTCGCCTGGCCTCACCCTG 116 782 1048097 N/A N/A  7056  7075ACTGTGACCCATGGATGCGG  98 783 1048113 N/A N/A  7343  7362CGCCCGTCCCCGTCCTGCCC  92 784 1048129 N/A N/A  7835  7854TGAAAAAGACTCAGTCCCTG 103 785 1048145 N/A N/A  7947  7966ATTGGTTGAATCCATCCATC 106 786 1048161 N/A N/A  8199  8218TCCTAGCTTTTTCCCCAGCA  92 787 1048177 N/A N/A  9318  9337CACTAATATTTAACATTAAG 120 788 1048193 N/A N/A  9475  9494GACATGCATATCTAGTGGTC  64 789 1048209 N/A N/A  9595  9614CTTTCCTACTTCTCTCTGCC 107 790 1048225 N/A N/A  9634  9653TGCTCAATACACATAGGTCC  95 791 1048241 N/A N/A  9806  9825GAAGTCCCAACTTAGCCACC 101 792 1048257 N/A N/A 10579 10598CCTGGCTTTCTGAAAACCCA  70 793 1048273 N/A N/A 10683 10702GGCTGGAGAGCCCCCAAATC 144 794 1048289 N/A N/A 10766 10785GGCTTCCCCATCGCACCCCC 117 795 1048305 N/A N/A 10930 10949ATCCCTCCTTACCTCTCCAT 106 796 1048321 N/A N/A 10998 11017CCCCAGGCTTTCCTCCATGG  81 797 1048337 N/A N/A 11113 11132CCTTAGAACAGCCTATGGAG 162 798 1048353 N/A N/A 11305 11324TGGTAGTTTCCTTTTACCAA  89 799 1048369 N/A N/A 11339 11358GGCGGATACGCCAAATCCCC  93 800 1048385 N/A N/A 11547 11566ACAGACCCCAGGTCCACCAC 104 801

TABLE 12Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047154  120  139  3568  3587GGCCCCCCACCATCATCTCC  97 802 1047170  226  245  3674  3693GCATTGAGTGCCCCAGCCAG 128 803 1047186  471  490  3919  3938CCCTCTCAACCTCCAGCCGG   3* 804 1047202  541  560  4926  4945GCTTCCAGCCTCAGGTTGGT  18* 805 1047218  628  647  5217  5236TCCAGCGACTCAATCTTCCT 100 806 1047234  723  742  5609  5628CGTCAAGCTCCACATGGACC 144 807 1047250 1017 1036  7589  7608AACTGGCCGCCTCCCGCACG 127 808 1047266 1206 1225  8355  8374GGTTGGAGAAGGTCTGCACG  77 809 1047282 1275 1294 10889 10908CCACGATGTTCCTCTTGAGG 157 810 1047298 1322 1341 11611 11630CTTGGACTCCTTAATGACCT  64 811 1047314 1395 1414 11684 11703GCTCGGGCCCCTCATGAGAC 100 812 1047330 1606 1625 11895 11914CATGCCCCTCCAGACTGCCC 106 813 1047346 1657 1676 11946 11965CGCCCTCCTCCCCTTCTCTC  79 814 1047362 1718 1737 12007 12026GGCAACAGTTTCCATAACAA  31 815 1047378 1771 1790 12060 12079TGAGGAAACTCAAAGGCACA  79 816 1047394 1835 1854 12124 12143CAGGGCTACCTTGTCTGTGG  40 817 1047410 2025 2044 12314 12333ATCTTAGACTGATCAGGGTC  67 818 1047426 2167 2186 12456 12475AGCTCCCACCTGCCCACAGC  94 819 1047442 2226 2245 12515 12534CCTCCCAGCACCTCATCCCT  97 820 1047458 2268 2287 12557 12576TCTCTGTACCCACAGCTGGG  84 821 1047474 2371 2390 12660 12679CACAACCCCTACTTGTATGC  61 822 1047490 2442 2461 12731 12750TTCAGCATCTTCAAGAGGAT  92 823 1047506 2470 2489 12759 12778CCATGGATACATCCCCTTTC 151 824 1047522 2749 2768 13038 13057CCAGTGTCTTCACTTTGCTC  44 825 1047538 2847 2866 13136 13155CAGTCTGCTCACCAGTCTGC 103 826 1047554 2885 2904 13174 13193TGGGCCCTCCCAGTCCCATC 172 827 1047570 2958 2977 13247 13266GGAAAGGACACCAAGTCTTG  90 828 1047586 3051 3070 13340 13359ATTTGTCTTTATTTTTCCTC  32 829 1047602 N/A N/A  8754  8773TGTGATTTTCCCCGTCTTTG  54 830 1047618 N/A N/A  8902  8921GGTGAGGCTCACTCCCTGTC  65 831 1047634 N/A N/A  9061  9080CACCACCTTTACCACTAACA  88 832 1047650 N/A N/A  9109  9128CTGCTTGCTCAGAGGCCCCA 115 833 1047666 N/A N/A  8372  8391CTGACCTCGAATCTGCAGGT  84 834 1047682 N/A N/A  8412  8431GGGCAGGACTCCAGTGCCCT 118 835 1047698 N/A N/A  8474  8493CTGGATCCCTTTGCCCTGAT 100 836 1047714 N/A N/A  8626  8645CACCCAGTTCTGCTGTCGAA  94 837 1047730 N/A N/A  8696  8715CAAAAACAGAAAACACTCAG 160 838 1047746 N/A N/A  3987  4006ACAAGGCCCCCCTTCCCCAT  57 839 1047762 N/A N/A  4215  4234CACTGCTTTCCCCAGTAGGG  48 840 1047778 N/A N/A  4314  4333CTCCTCCTTTATATGGACAC  96 841 1047794 N/A N/A  4345  4364CCTCATGGACATCAACCTTC  92 842 1047810 N/A N/A  4379  4398TTCCCTCACTTTCTTCACCC 102 843 1047826 N/A N/A  4515  4534TCCCAGGTAACCACCTTTTG  76 844 1047842 N/A N/A  4691  4710CTAAGGTGCCTTATCAGGGT  78 845 1047858 N/A N/A  4794  4813ACCCAGGACCAGTAGAGCAG  84 846 1047874 N/A N/A  4843  4862AATACCTGCCTCAGTCTCCC 106 847 1047890 N/A N/A  4999  5018CAATCCTTTCCTCCCTCCCC 109 848 1047906 N/A N/A  5104  5123CTCCCCATTCCTCAGCCTTG  90 849 1047922 N/A N/A  5334  5353CTCAGCTTCTCTGTCTCTGT 123 850 1047938 N/A N/A  5465  5484CCCCTCGGCCAGGAGTTCGA 123 851 1047954 N/A N/A  5781  5800CCCCATTCTCTTGTACAGAG  98 852 1047970 N/A N/A  5817  5836GAGAGGATATTCTCCCAGCT 151 853 1047986 N/A N/A  6001  6020AAGAACAGCACCTACTTCAT  85 854 1048002 N/A N/A  6090  6109GAGTTGGAAATCCAACTCTA  97 855 1048018 N/A N/A  6159  6178GTAGAGGCTCAGTAACCCAA  51 856 1048034 N/A N/A  6310  6329ATGCCCCCTCTACAGTGTCT  71 857 1048050 N/A N/A  6366  6385AATGGCCCTCCCTTCTTCTC 177 858 1048066 N/A N/A  6503  6522CCATCGGGCCCTCACCCTGC 110 859 1048082 N/A N/A  6951  6970ACCTTCCACACTGACAGCTG 169 860 1048098 N/A N/A  7058  7077CAACTGTGACCCATGGATGC  78 861 1048114 N/A N/A  7349  7368CTGCTCCGCCCGTCCCCGTC 120 862 1048130 N/A N/A  7836  7855CTGAAAAAGACTCAGTCCCT 118 863 1048146 N/A N/A  7949  7968ATATTGGTTGAATCCATCCA 102 864 1048162 N/A N/A  8211  8230TCTAACTCCATCTCCTAGCT  98 865 1048178 N/A N/A  9319  9338TCACTAATATTTAACATTAA  84 866 1048194 N/A N/A  9492  9511GCTGAATTAAGTCCTGAGAC  55 867 1048210 N/A N/A  9596  9615TCTTTCCTACTTCTCTCTGC  82 868 1048226 N/A N/A  9662  9681AGGCTGTTAAACATGTGGCA  71 869 1048242 N/A N/A  9807  9826AGAAGTCCCAACTTAGCCAC  85 870 1048258 N/A N/A 10580 10599ACCTGGCTTTCTGAAAACCC  79 871 1048274 N/A N/A 10698 10717AGGCTCTTCCAAACGGGCTG 103 872 1048290 N/A N/A 10768 10787CCGGCTTCCCCATCGCACCC  81 873 1048306 N/A N/A 10931 10950AATCCCTCCTTACCTCTCCA 116 874 1048322 N/A N/A 11004 11023TGCCAGCCCCAGGCTTTCCT  66 875 1048338 N/A N/A 11117 11136CTCCCCTTAGAACAGCCTAT  88 876 1048354 N/A N/A 11306 11325CTGGTAGTTTCCTTTTACCA 113 877 1048370 N/A N/A 11340 11359TGGCGGATACGCCAAATCCC  75 878 1048386 N/A N/A 11555 11574GAGTTCACACAGACCCCAGG  85 879

TABLE 13Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047155  121  140  3569  3588AGGCCCCCCACCATCATCTC 124 880 1047171  228  247  3676  3695CAGCATTGAGTGCCCCAGCC 104 881 1047187  472  491  3920  3939TCCCTCTCAACCTCCAGCCG   7* 882 1047203  542  561  4927  4946GGCTTCCAGCCTCAGGTTGG  18* 883 1047219  629  648  5218  5237CTCCAGCGACTCAATCTTCC  86 884 1047235  761  780  5647  5666GATCTCTTTCAGGGCTGCGG  51 885 1047251 1019 1038  7591  7610ATAACTGGCCGCCTCCCGCA 143 886 1047267 1223 1242 N/A N/AGGTTTCTCGAATCTGCAGGT  65 887 1047283 1280 1299 10894 10913CTTCACCACGATGTTCCTCT  73 888 1047299 1323 1342 11612 11631GCTTGGACTCCTTAATGACC  91 889 1047315 1396 1415 11685 11704TGCTCGGGCCCCTCATGAGA  84 890 1047331 1607 1626 11896 11915CCATGCCCCTCCAGACTGCC  66 891 1047347 1658 1677 11947 11966CCGCCCTCCTCCCCTTCTCT  73 892 1047363 1721 1740 12010 12029TCTGGCAACAGTTTCCATAA  66 893 1047379 1772 1791 12061 12080CTGAGGAAACTCAAAGGCAC 100 894 1047395 1837 1856 12126 12145GCCAGGGCTACCTTGTCTGT  80 895 1047411 2026 2045 12315 12334CATCTTAGACTGATCAGGGT  72 896 1047427 2169 2188 12458 12477CAAGCTCCCACCTGCCCACA 111 897 1047443 2232 2251 12521 12540TCTATCCCTCCCAGCACCTC  80 898 1047459 2269 2288 12558 12577CTCTCTGTACCCACAGCTGG  85 899 1047475 2372 2391 12661 12680CCACAACCCCTACTTGTATG  69 900 1047491 2443 2462 12732 12751TTTCAGCATCTTCAAGAGGA 121 901 1047507 2471 2490 12760 12779CCCATGGATACATCCCCTTT  79 902 1047523 2751 2770 13040 13059AGCCAGTGTCTTCACTTTGC  60 903 1047539 2855 2874 13144 13163GATCCCACCAGTCTGCTCAC  84 904 1047555 2886 2905 13175 13194GTGGGCCCTCCCAGTCCCAT  81 905 1047571 3010 3029 13299 13318TGCCCTGAAGATTAGCAGCA 103 906 1047587 3052 3071 13341 13360CATTTGTCTTTATTTTTCCT  14 907 1047603 N/A N/A  8755  8774TTGTGATTTTCCCCGTCTTT  75 908 1047619 N/A N/A  8969  8988ACGCAGTCCAGGCCCTTTAG  57 909 1047635 N/A N/A  9063  9082CTCACCACCTTTACCACTAA 113 910 1047651 N/A N/A  9128  9147AGAGGTGAGACAGAGGCTGC 112 911 1047667 N/A N/A  8374  8393TACTGACCTCGAATCTGCAG  85 912 1047683 N/A N/A  8432  8451CCTACAGGCCCTGGAGGAGG  85 913 1047699 N/A N/A  8476  8495AGCTGGATCCCTTTGCCCTG  97 914 1047715 N/A N/A  8630  8649CTCTCACCCAGTTCTGCTGT  77 915 1047731 N/A N/A  8723  8742CCCTGTAGTGACAAGCAGTT  84 916 1047747 N/A N/A  3997  4016CCTTCTGCTCACAAGGCCCC  93 917 1047763 N/A N/A  4257  4276AAGCCCAGCCATGAATGAAA  83 918 1047779 N/A N/A  4316  4335AACTCCTCCTTTATATGGAC  91 919 1047795 N/A N/A  4353  4372CCAATCTCCCTCATGGACAT  66 920 1047811 N/A N/A  4386  4405CTGCTCTTTCCCTCACTTTC 103 921 1047827 N/A N/A  4516  4535ATCCCAGGTAACCACCTTTT  85 922 1047843 N/A N/A  4692  4711ACTAAGGTGCCTTATCAGGG 117 923 1047859 N/A N/A  4795  4814CACCCAGGACCAGTAGAGCA  56 924 1047875 N/A N/A  4848  4867ACTTGAATACCTGCCTCAGT  85 925 1047891 N/A N/A  5001  5020ATCAATCCTTTCCTCCCTCC 102 926 1047907 N/A N/A  5107  5126CTTCTCCCCATTCCTCAGCC  56 927 1047923 N/A N/A  5349  5368AGTGTCTCTCTCAGTCTCAG  83 928 1047939 N/A N/A  5477  5496CTCTTCTGCCTGCCCCTCGG 101 929 1047955 N/A N/A  5783  5802TCCCCCATTCTCTTGTACAG  49 930 1047971 N/A N/A  5818  5837GGAGAGGATATTCTCCCAGC  93 931 1047987 N/A N/A  6007  6026CTGTCAAAGAACAGCACCTA 112 932 1048003 N/A N/A  6091  6110AGAGTTGGAAATCCAACTCT  92 933 1048019 N/A N/A  6201  6220GGTCAGACACCTCTCTGTGT  82 934 1048035 N/A N/A  6321  6340GCCTAGCCCAAATGCCCCCT  71 935 1048051 N/A N/A  6384  6403GCTCTGTCCTCCACTAGGAA  97 936 1048067 N/A N/A  6504  6523CCCATCGGGCCCTCACCCTG  88 937 1048083 N/A N/A  6954  6973AGAACCTTCCACACTGACAG  76 938 1048099 N/A N/A  7060  7079AACAACTGTGACCCATGGAT  74 939 1048115 N/A N/A  7351  7370CCCTGCTCCGCCCGTCCCCG  98 940 1048131 N/A N/A  7837  7856GCTGAAAAAGACTCAGTCCC  84 941 1048147 N/A N/A  7973  7992GTCCTTGGCCTTGAGGCCTA  95 942 1048163 N/A N/A  8213  8232AGTCTAACTCCATCTCCTAG  88 943 1048179 N/A N/A  9322  9341CATTCACTAATATTTAACAT 110 944 1048195 N/A N/A  9507  9526GTTAGCCTTTCTGATGCTGA  53 945 1048211 N/A N/A  9597  9616CTCTTTCCTACTTCTCTCTG  91 946 1048227 N/A N/A  9708  9727GGCCTACTTCTCTAGGTGGG  82 947 1048243 N/A N/A  9808  9827AAGAAGTCCCAACTTAGCCA 141 948 1048259 N/A N/A 10602 10621CCTCTGCAAGCCCTGGCCTG  95 949 1048275 N/A N/A 10699 10718TAGGCTCTTCCAAACGGGCT  89 950 1048291 N/A N/A 10769 10788CCCGGCTTCCCCATCGCACC  79 951 1048307 N/A N/A 10934 10953CCAAATCCCTCCTTACCTCT 156 952 1048323 N/A N/A 11017 11036GGAACCTTCTATGTGCCAGC  74 953 1048339 N/A N/A 11132 11151GCTTTGGTACCAAGGCTCCC 192 954 1048355 N/A N/A 11307 11326CCTGGTAGTTTCCTTTTACC  52 955 1048371 N/A N/A 11379 11398GCTGGAGTAAGATGAGCTCC  86 956 1048387 N/A N/A 11570 11589CAGTGCAACAGTTAGGAGTT  66 957

TABLE 14Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047156  122  141  3570  3589CAGGCCCCCCACCATCATCT  90  958 1047172  237  256  3685  3704CCTTGAAGCCAGCATTGAGT  89  959 1047188  473  492  3921  3940GTCCCTCTCAACCTCCAGCC   6*  960 1047204  545  564  4930  4949CTCGGCTTCCAGCCTCAGGT  37*  961 1047220  634  653  5223  5242TCCTCCTCCAGCGACTCAAT  94  962 1047236  766  785  5652  5671GTGCGGATCTCTTTCAGGGC  63  963 1047252 1020 1039  7592  7611GATAACTGGCCGCCTCCCGC 113  964 1047268 1225 1244 N/A N/ACTGGTTTCTCGAATCTGCAG  83  965 1047284 1283 1302 10897 10916GGTCTTCACCACGATGTTCC  94  966 1047300 1345 1364 11634 11653CACATCACATCCTTGTGCTC  75  967 1047316 1398 1417 11687 11706TCTGCTCGGGCCCCTCATGA  96  968 1047332 1608 1627 11897 11916GCCATGCCCCTCCAGACTGC  73  969 1047348 1659 1678 11948 11967CCCGCCCTCCTCCCCTTCTC 107  970 1047364 1723 1742 12012 12031TCTCTGGCAACAGTTTCCAT  65  971 1047380 1773 1792 12062 12081CCTGAGGAAACTCAAAGGCA  92  972 1047396 1838 1857 12127 12146GGCCAGGGCTACCTTGTCTG  67  973 1047412 2027 2046 12316 12335CCATCTTAGACTGATCAGGG  76  974 1047428 2170 2189 12459 12478TCAAGCTCCCACCTGCCCAC 127  975 1047444 2233 2252 12522 12541CTCTATCCCTCCCAGCACCT  42  976 1047460 2270 2289 12559 12578CCTCTCTGTACCCACAGCTG  94  977 1047476 2373 2392 12662 12681CCCACAACCCCTACTTGTAT 115  978 1047492 2449 2468 12738 12757CTCCTGTTTCAGCATCTTCA  45  979 1047508 2472 2491 12761 12780CCCCATGGATACATCCCCTT  86  980 1047524 2759 2778 13048 13067GGAATATGAGCCAGTGTCTT  63  981 1047540 2857 2876 13146 13165CAGATCCCACCAGTCTGCTC  89  982 1047556 2889 2908 13178 13197GAAGTGGGCCCTCCCAGTCC 113  983 1047572 3011 3030 13300 13319GTGCCCTGAAGATTAGCAGC  85  984 1047588 3053 3072 13342 13361GCATTTGTCTTTATTTTTCC  11  985 1047604 N/A N/A  8756  8775CTTGTGATTTTCCCCGTCTT  74  986 1047620 N/A N/A  8972  8991ATGACGCAGTCCAGGCCCTT  78  987 1047636 N/A N/A  9064  9083ACTCACCACCTTTACCACTA  85  988 1047652 N/A N/A  9129  9148AAGAGGTGAGACAGAGGCTG  66  989 1047668 N/A N/A  8375  8394GTACTGACCTCGAATCTGCA  96  990 1047684 N/A N/A  8437  8456AGCAACCTACAGGCCCTGGA 116  991 1047700 N/A N/A  8477  8496GAGCTGGATCCCTTTGCCCT  82  992 1047716 N/A N/A  8631  8650GCTCTCACCCAGTTCTGCTG  83  993 1047732 N/A N/A  8725  8744CCCCCTGTAGTGACAAGCAG  71  994 1047748 N/A N/A  4051  4070GAGGTTCGGCCCCTCCCTGA  68  995 1047764 N/A N/A  4258  4277AAAGCCCAGCCATGAATGAA 111  996 1047780 N/A N/A  4317  4336CAACTCCTCCTTTATATGGA 104  997 1047796 N/A N/A  4355  4374ATCCAATCTCCCTCATGGAC  78  998 1047812 N/A N/A  4388  4407GCCTGCTCTTTCCCTCACTT  98  999 1047828 N/A N/A  4520  4539TCTGATCCCAGGTAACCACC  81 1000 1047844 N/A N/A  4693  4712TACTAAGGTGCCTTATCAGG  92 1001 1047860 N/A N/A  4800  4819ACCCCCACCCAGGACCAGTA  94 1002 1047876 N/A N/A  4851  4870GACACTTGAATACCTGCCTC  89 1003 1047892 N/A N/A  5002  5021CATCAATCCTTTCCTCCCTC  79 1004 1047908 N/A N/A  5109  5128TCCTTCTCCCCATTCCTCAG  88 1005 1047924 N/A N/A  5355  5374TCTCTGAGTGTCTCTCTCAG  92 1006 1047940 N/A N/A  5478  5497CCTCTTCTGCCTGCCCCTCG 111 1007 1047956 N/A N/A  5784  5803TTCCCCCATTCTCTTGTACA  98 1008 1047972 N/A N/A  5836  5855GGTGAAAGTCAGTCACCTGG  90 1009 1047988 N/A N/A  6009  6028ATCTGTCAAAGAACAGCACC  96 1010 1048004 N/A N/A  6092  6111TAGAGTTGGAAATCCAACTC  92 1011 1048020 N/A N/A  6211  6230ACACCTTCCAGGTCAGACAC  71 1012 1048036 N/A N/A  6322  6341TGCCTAGCCCAAATGCCCCC 111 1013 1048052 N/A N/A  6386  6405AGGCTCTGTCCTCCACTAGG  87 1014 1048068 N/A N/A  6513  6532CCTCCCAGCCCCATCGGGCC 146 1015 1048084 N/A N/A  6955  6974CAGAACCTTCCACACTGACA 123 1016 1048100 N/A N/A  7065  7084TTCCCAACAACTGTGACCCA  69 1017 1048116 N/A N/A  7387  7406TGGCCCTTCTCCCCTGGCAT 122 1018 1048132 N/A N/A  7840  7859AAGGCTGAAAAAGACTCAGT  82 1019 1048148 N/A N/A  7982  8001GTGACCCAAGTCCTTGGCCT  85 1020 1048164 N/A N/A  8217  8236GGAAAGTCTAACTCCATCTC  93 1021 1048180 N/A N/A  9323  9342ACATTCACTAATATTTAACA  91 1022 1048196 N/A N/A  9510  9529CTGGTTAGCCTTTCTGATGC  51 1023 1048212 N/A N/A  9599  9618TCCTCTTTCCTACTTCTCTC  73 1024 1048228 N/A N/A  9709  9728GGGCCTACTTCTCTAGGTGG  48 1025 1048244 N/A N/A  9813  9832AGCTCAAGAAGTCCCAACTT  87 1026 1048260 N/A N/A 10616 10635GCTTCATTTCAGCCCCTCTG  82 1027 1048276 N/A N/A 10700 10719CTAGGCTCTTCCAAACGGGC  94 1028 1048292 N/A N/A 10771 10790TGCCCGGCTTCCCCATCGCA  98 1029 1048308 N/A N/A 10936 10955GCCCAAATCCCTCCTTACCT  95 1030 1048324 N/A N/A 11021 11040GCTGGGAACCTTCTATGTGC  64 1031 1048340 N/A N/A 11133 11152GGCTTTGGTACCAAGGCTCC  89 1032 1048356 N/A N/A 11309 11328CCCCTGGTAGTTTCCTTTTA  94 1033 1048372 N/A N/A 11433 11452TGGTGAGATAACACTGGGAA  51 1034 1048388 N/A N/A 11571 11590ACAGTGCAACAGTTAGGAGT  73 1035

TABLE 15Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047157  123  142  3571  3590CCAGGCCCCCCACCATCATC 116 1036 1047173  239  258  3687  3706CTCCTTGAAGCCAGCATTGA  95 1037 1047189  474  493  3922  3941TGTCCCTCTCAACCTCCAGC  11* 1038 1047205  547  566  4932  4951TTCTCGGCTTCCAGCCTCAG  31* 1039 1047221  637  656  5226  5245ATCTCCTCCTCCAGCGACTC  83 1040 1047237  786  805  5672  5691ACGCCATTGCCTCATACTGC 104 1041 1047253 1021 1040  7593  7612TGATAACTGGCCGCCTCCCG  94 1042 1047269 1227 1246 N/A N/AGGCTGGTTTCTCGAATCTGC  92 1043 1047285 1285 1304 10899 10918ACGGTCTTCACCACGATGTT  86 1044 1047301 1346 1365 11635 11654TCACATCACATCCTTGTGCT  79 1045 1047317 1406 1425 11695 11714ATCCTGCTTCTGCTCGGGCC  98 1046 1047333 1610 1629 11899 11918TGGCCATGCCCCTCCAGACT  74 1047 1047349 1660 1679 11949 11968CCCCGCCCTCCTCCCCTTCT  98 1048 1047365 1724 1743 12013 12032ATCTCTGGCAACAGTTTCCA  60 1049 1047381 1774 1793 12063 12082GCCTGAGGAAACTCAAAGGC  91 1050 1047397 1839 1858 12128 12147TGGCCAGGGCTACCTTGTCT  82 1051 1047413 2033 2052 12322 12341CCCCACCCATCTTAGACTGA  84 1052 1047429 2172 2191 12461 12480AATCAAGCTCCCACCTGCCC  45 1053 1047445 2234 2253 12523 12542CCTCTATCCCTCCCAGCACC 104 1054 1047461 2275 2294 12564 12583CTTGACCTCTCTGTACCCAC  67 1055 1047477 2375 2394 12664 12683CACCCACAACCCCTACTTGT  97 1056 1047493 2450 2469 12739 12758TCTCCTGTTTCAGCATCTTC  58 1057 1047509 2478 2497 12767 12786CCCTGCCCCCATGGATACAT  76 1058 1047525 2765 2784 13054 13073GCTGCAGGAATATGAGCCAG  84 1059 1047541 2858 2877 13147 13166ACAGATCCCACCAGTCTGCT  98 1060 1047557 2890 2909 13179 13198TGAAGTGGGCCCTCCCAGTC  87 1061 1047573 3016 3035 13305 13324CAGCAGTGCCCTGAAGATTA  45 1062 1047589 3054 3073 13343 13362AGCATTTGTCTTTATTTTTC  17 1063 1047605 N/A N/A  8757  8776CCTTGTGATTTTCCCCGTCT  64 1064 1047621 N/A N/A  9023  9042TACAGTTACTCTGTACCACG 118 1065 1047637 N/A N/A  9065  9084GACTCACCACCTTTACCACT  87 1066 1047653 N/A N/A  9199  9218GATGAAAGAATAAAGCAGAG 102 1067 1047669 N/A N/A  8376  8395TGTACTGACCTCGAATCTGC  90 1068 1047685 N/A N/A  8439  8458GGAGCAACCTACAGGCCCTG 104 1069 1047701 N/A N/A  8478  8497AGAGCTGGATCCCTTTGCCC  87 1070 1047717 N/A N/A  8632  8651AGCTCTCACCCAGTTCTGCT  62 1071 1047733 N/A N/A  8731  8750CTTTTGCCCCCTGTAGTGAC  56 1072 1047749 N/A N/A  4088  4107GTGCCCCATCAAGAGGTAGG 147 1073 1047765 N/A N/A  4260  4279ACAAAGCCCAGCCATGAATG 142 1074 1047781 N/A N/A  4318  4337CCAACTCCTCCTTTATATGG  74 1075 1047797 N/A N/A  4356  4375AATCCAATCTCCCTCATGGA  93 1076 1047813 N/A N/A  4390  4409CTGCCTGCTCTTTCCCTCAC  99 1077 1047829 N/A N/A  4521  4540CTCTGATCCCAGGTAACCAC  96 1078 1047845 N/A N/A  4700  4719AACTCATTACTAAGGTGCCT 101 1079 1047861 N/A N/A  4801  4820CACCCCCACCCAGGACCAGT  90 1080 1047877 N/A N/A  4852  4871GGACACTTGAATACCTGCCT  84 1081 1047893 N/A N/A  5003  5022CCATCAATCCTTTCCTCCCT  82 1082 1047909 N/A N/A  5112  5131GGCTCCTTCTCCCCATTCCT  92 1083 1047925 N/A N/A  5368  5387TGTTTCTCTCCTCTCTCTGA 113 1084 1047941 N/A N/A  5484  5503TTGTGTCCTCTTCTGCCTGC  93 1085 1047957 N/A N/A  5786  5805CCTTCCCCCATTCTCTTGTA  87 1086 1047973 N/A N/A  5842  5861TTCTCTGGTGAAAGTCAGTC  95 1087 1047989 N/A N/A  6012  6031CTCATCTGTCAAAGAACAGC 102 1088 1048005 N/A N/A  6110  6129CTCCCAAGTGAGATGTGCTA 112 1089 1048021 N/A N/A  6212  6231CACACCTTCCAGGTCAGACA  70 1090 1048037 N/A N/A  6323  6342CTGCCTAGCCCAAATGCCCC  86 1091 1048053 N/A N/A  6396  6415TTCTGCCTCCAGGCTCTGTC 117 1092 1048069 N/A N/A  6519  6538GGAGGTCCTCCCAGCCCCAT  94 1093 1048085 N/A N/A  6956  6975CCAGAACCTTCCACACTGAC  79 1094 1048101 N/A N/A  7069  7088ACTTTTCCCAACAACTGTGA  84 1095 1048117 N/A N/A  7388  7407CTGGCCCTTCTCCCCTGGCA  81 1096 1048133 N/A N/A  7841  7860CAAGGCTGAAAAAGACTCAG 121 1097 1048149 N/A N/A  7985  8004AGGGTGACCCAAGTCCTTGG  89 1098 1048165 N/A N/A  8219  8238CAGGAAAGTCTAACTCCATC  80 1099 1048181 N/A N/A  9324  9343CACATTCACTAATATTTAAC  93   21 1048197 N/A N/A  9512  9531GCCTGGTTAGCCTTTCTGAT  59 1100 1048213 N/A N/A  9604  9623CCCTTTCCTCTTTCCTACTT  71 1101 1048229 N/A N/A  9727  9746TGTAAAATAAGGATGATGGG 101 1102 1048245 N/A N/A  9816  9835CAGAGCTCAAGAAGTCCCAA 101 1103 1048261 N/A N/A 10617 10636GGCTTCATTTCAGCCCCTCT  73 1104 1048277 N/A N/A 10702 10721GCCTAGGCTCTTCCAAACGG 105 1105 1048293 N/A N/A 10784 10803CCATCCTCTCCCATGCCCGG 100 1106 1048309 N/A N/A 10937 10956GGCCCAAATCCCTCCTTACC 118 1107 1048325 N/A N/A 11058 11077CTCTCCTCCAGAATTCCCTG  76 1108 1048341 N/A N/A 11149 11168TAGGATCCCATCTAGTGGCT  51 1109 1048357 N/A N/A 11316 11335TCCCATTCCCCTGGTAGTTT  57 1110 1048373 N/A N/A 11435 11454GGTGGTGAGATAACACTGGG  88 1111 1048389 N/A N/A 11608 11627GGACTCCTTAATGACCTGCA  53 1112

TABLE 16Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047158  124  143  3572  3591GCCAGGCCCCCCACCATCAT 102 1113 1047174  240  259  3688  3707TCTCCTTGAAGCCAGCATTG 109 1114 1047190  477  496  3925  3944GATTGTCCCTCTCAACCTCC  17* 1115 1047206  563  582  4948  4967ATAGGCAGCCAGGTTGTTCT  35* 1116 1047222  638  657  5227  5246GATCTCCTCCTCCAGCGACT  75 1117 1047238  809  828  5695  5714TTCGGCTTCATGCATGTTGC 123 1118 1047254 1026 1045  7598  7617CCTCCTGATAACTGGCCGCC 117 1119 1047270 1228 1247 N/A N/AAGGCTGGTTTCTCGAATCTG 117 1120 1047286 1286 1305 10900 10919CACGGTCTTCACCACGATGT  89 1121 1047302 1348 1367 11637 11656CCTCACATCACATCCTTGTG 103 1122 1047318 1413 1432 11702 11721AGCAACTATCCTGCTTCTGC  58 1123 1047334 1612 1631 11901 11920GCTGGCCATGCCCCTCCAGA 114 1124 1047350 1661 1680 11950 11969CCCCCGCCCTCCTCCCCTTC  90 1125 1047366 1731 1750 12020 12039AACCTCCATCTCTGGCAACA  59 1126 1047382 1782 1801 12071 12090CTCCAGCAGCCTGAGGAAAC 131 1127 1047398 1844 1863 12133 12152GCCTCTGGCCAGGGCTACCT 100 1128 1047414 2034 2053 12323 12342TCCCCACCCATCTTAGACTG 113 1129 1047430 2178 2197 12467 12486GCTGAGAATCAAGCTCCCAC  54 1130 1047446 2235 2254 12524 12543CCCTCTATCCCTCCCAGCAC 100 1131 1047462 2278 2297 12567 12586GGGCTTGACCTCTCTGTACC  73 1132 1047478 2376 2395 12665 12684TCACCCACAACCCCTACTTG  81 1133 1047494 2451 2470 12740 12759CTCTCCTGTTTCAGCATCTT  61 1134 1047510 2481 2500 12770 12789ATGCCCTGCCCCCATGGATA  83 1135 1047526 2777 2796 13066 13085CCCGGCCTCCAGGCTGCAGG  89 1136 1047542 2859 2878 13148 13167CACAGATCCCACCAGTCTGC  85 1137 1047558 2901 2920 13190 13209GAGGAGAACCCTGAAGTGGG  95 1138 1047574 3018 3037 13307 13326AGCAGCAGTGCCCTGAAGAT  78 1139 1047590 3055 3074 13344 13363CAGCATTTGTCTTTATTTTT  21 1140 1047606 N/A N/A  8758  8777ACCTTGTGATTTTCCCCGTC  70 1141 1047622 N/A N/A  9024  9043GTACAGTTACTCTGTACCAC 138 1142 1047638 N/A N/A  9066  9085GGACTCACCACCTTTACCAC  68 1143 1047654 N/A N/A  9200  9219GGATGAAAGAATAAAGCAGA  85 1144 1047670 N/A N/A  8377  8396CTGTACTGACCTCGAATCTG  93 1145 1047686 N/A N/A  8443  8462GTCTGGAGCAACCTACAGGC  79 1146 1047702 N/A N/A  8495  8514CACGAAGGCCCCCAGGGAGA  77 1147 1047718 N/A N/A  8633  8652AAGCTCTCACCCAGTTCTGC 108 1148 1047734 N/A N/A  8733  8752TGCTTTTGCCCCCTGTAGTG  47 1149 1047750 N/A N/A  4090  4109TAGTGCCCCATCAAGAGGTA  88 1150 1047766 N/A N/A  4262  4281TCACAAAGCCCAGCCATGAA  89 1151 1047782 N/A N/A  4319  4338TCCAACTCCTCCTTTATATG  85 1152 1047798 N/A N/A  4357  4376GAATCCAATCTCCCTCATGG  94 1153 1047814 N/A N/A  4396  4415CCAGACCTGCCTGCTCTTTC  89 1154 1047830 N/A N/A  4523  4542TCCTCTGATCCCAGGTAACC  76 1155 1047846 N/A N/A  4701  4720TAACTCATTACTAAGGTGCC  79 1156 1047862 N/A N/A  4804  4823GACCACCCCCACCCAGGACC 105 1157 1047878 N/A N/A  4853  4872AGGACACTTGAATACCTGCC  56 1158 1047894 N/A N/A  5004  5023GCCATCAATCCTTTCCTCCC 100 1159 1047910 N/A N/A  5113  5132AGGCTCCTTCTCCCCATTCC  90 1160 1047926 N/A N/A  5379  5398CTGCCAATCTCTGTTTCTCT  85 1161 1047942 N/A N/A  5496  5515TTCCCCACGCCATTGTGTCC  76 1162 1047958 N/A N/A  5787  5806TCCTTCCCCCATTCTCTTGT 121 1163 1047974 N/A N/A  5851  5870CCATCTCACTTCTCTGGTGA  82 1164 1047990 N/A N/A  6019  6038CGGCTCTCTCATCTGTCAAA  43 1165 1048006 N/A N/A  6115  6134GCAGGCTCCCAAGTGAGATG  99 1166 1048022 N/A N/A  6221  6240CGTCAATATCACACCTTCCA  96 1167 1048038 N/A N/A  6324  6343CCTGCCTAGCCCAAATGCCC 119 1168 1048054 N/A N/A  6400  6419GCTTTTCTGCCTCCAGGCTC  77 1169 1048070 N/A N/A  6539  6558CCTTTCTCCCCTGCCTGCAG  89 1170 1048086 N/A N/A  6957  6976TCCAGAACCTTCCACACTGA  82 1171 1048102 N/A N/A  7070  7089GACTTTTCCCAACAACTGTG  84 1172 1048118 N/A N/A  7390  7409CCCTGGCCCTTCTCCCCTGG  76 1173 1048134 N/A N/A  7842  7861ACAAGGCTGAAAAAGACTCA  83 1174 1048150 N/A N/A  7986  8005GAGGGTGACCCAAGTCCTTG 110 1175 1048166 N/A N/A  8221  8240GCCAGGAAAGTCTAACTCCA  75 1176 1048182 N/A N/A  9326  9345GTCACATTCACTAATATTTA  44 1177 1048198 N/A N/A  9525  9544CCTCTACTAGTCAGCCTGGT  65 1178 1048214 N/A N/A  9606  9625TCCCCTTTCCTCTTTCCTAC  75 1179 1048230 N/A N/A  9772  9791CTCTGGGCAAGTTAATTGAC 122 1180 1048246 N/A N/A  9818  9837ACCAGAGCTCAAGAAGTCCC  83 1181 1048262 N/A N/A 10618 10637TGGCTTCATTTCAGCCCCTC  81 1182 1048278 N/A N/A 10710 10729CAGAGAGAGCCTAGGCTCTT  95 1183 1048294 N/A N/A 10788 10807TGAGCCATCCTCTCCCATGC 125 1184 1048310 N/A N/A 10940 10959CTGGGCCCAAATCCCTCCTT  83 1185 1048326 N/A N/A 11060 11079TGCTCTCCTCCAGAATTCCC 123 1186 1048342 N/A N/A 11223 11242CTAACTTTAATTCTCTTTCT 114 1187 1048358 N/A N/A 11317 11336TTCCCATTCCCCTGGTAGTT  89 1188 1048374 N/A N/A 11436 11455GGGTGGTGAGATAACACTGG 106 1189 1048390 N/A N/A 11609 11628TGGACTCCTTAATGACCTGC  68 1190

TABLE 17Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP RNA SEQ ID No. Site Site SiteSite Sequence (5′ to 3′) (% control) NO 1047159  126  145  3574  3593GAGCCAGGCCCCCCACCATC 104 1191 1047175  244  263  3692  3711CGGGTCTCCTTGAAGCCAGC 125 1192 1047191  480  499  3928  3947CCAGATTGTCCCTCTCAACC  15* 1193 1047207  574  593 N/A N/AGCTTCCTGTCTATAGGCAGC  37* 1194 1047223  639  658  5228  5247GGATCTCCTCCTCCAGCGAC  79 1195 1047239  811  830  5697  5716TCTTCGGCTTCATGCATGTT  98 1196 1047255 1028 1047  7600  7619CGCCTCCTGATAACTGGCCG 105 1197 1047271 1241 1260 10855 10874AGACTTGGTGTCCAGGCTGG  79 1198 1047287 1289 1308 10903 10922CTCCACGGTCTTCACCACGA 138 1199 1047303 1349 1368 11638 11657GCCTCACATCACATCCTTGT 113 1200 1047319 1414 1433 11703 11722GAGCAACTATCCTGCTTCTG  68 1201 1047335 1615 1634 11904 11923GCTGCTGGCCATGCCCCTCC 123 1202 1047351 1663 1682 11952 11971GCCCCCCGCCCTCCTCCCCT  73 1203 1047367 1734 1753 12023 12042GAGAACCTCCATCTCTGGCA  67 1204 1047383 1790 1809 12079 12098CAGTTTTCCTCCAGCAGCCT 109   11 1047399 1853 1872 12142 12161ACAAAACAAGCCTCTGGCCA  90 1206 1047415 2035 2054 12324 12343GTCCCCACCCATCTTAGACT  76 1207 1047431 2179 2198 12468 12487TGCTGAGAATCAAGCTCCCA  91 1208 1047447 2236 2255 12525 12544CCCCTCTATCCCTCCCAGCA  83 1209 1047463 2279 2298 12568 12587TGGGCTTGACCTCTCTGTAC 103 1210 1047479 2377 2396 12666 12685GTCACCCACAACCCCTACTT  88 1211 1047495 2456 2475 12745 12764CCTTTCTCTCCTGTTTCAGC  54 1212 1047511 2482 2501 12771 12790CATGCCCTGCCCCCATGGAT  83 1213 1047527 2779 2798 13068 13087CACCCGGCCTCCAGGCTGCA 106 1214 1047543 2860 2879 13149 13168GCACAGATCCCACCAGTCTG 108 1215 1047559 2902 2921 13191 13210AGAGGAGAACCCTGAAGTGG 120 1216 1047575 3034 3053 13323 13342CTCAGCGACTAAAGGCAGCA  59 1217 1047591 3056 3075 13345 13364GCAGCATTTGTCTTTATTTT  22 1218 1047607 N/A N/A  8760  8779TGACCTTGTGATTTTCCCCG  59 1219 1047623 N/A N/A  9025  9044TGTACAGTTACTCTGTACCA 103 1220 1047639 N/A N/A  9067  9086AGGACTCACCACCTTTACCA  77 1221 1047655 N/A N/A  9201  9220GGGATGAAAGAATAAAGCAG  67 1222 1047671 N/A N/A  8378  8397GCTGTACTGACCTCGAATCT  94 1223 1047687 N/A N/A  8453  8472CTCAGTCCCAGTCTGGAGCA 123 1224 1047703 N/A N/A  8502  8521GCAGTGTCACGAAGGCCCCC  89 1225 1047719 N/A N/A  8635  8654TCAAGCTCTCACCCAGTTCT  96 1226 1047735 N/A N/A  8735  8754GGTGCTTTTGCCCCCTGTAG  55 1227 1047751 N/A N/A  4091  4110ATAGTGCCCCATCAAGAGGT 135 1228 1047767 N/A N/A  4263  4282GTCACAAAGCCCAGCCATGA 122 1229 1047783 N/A N/A  4321  4340CTTCCAACTCCTCCTTTATA  95 1230 1047799 N/A N/A  4358  4377AGAATCCAATCTCCCTCATG  67 1231 1047815 N/A N/A  4399  4418CGCCCAGACCTGCCTGCTCT  94 1232 1047831 N/A N/A  4524  4543TTCCTCTGATCCCAGGTAAC  86 1233 1047847 N/A N/A  4702  4721TTAACTCATTACTAAGGTGC  87 1234 1047863 N/A N/A  4805  4824AGACCACCCCCACCCAGGAC  96 1235 1047879 N/A N/A  4866  4885CCAGGCTCTTCTGAGGACAC 119 1236 1047895 N/A N/A  5005  5024GGCCATCAATCCTTTCCTCC  81 1237 1047911 N/A N/A  5115  5134CCAGGCTCCTTCTCCCCATT  92 1238 1047927 N/A N/A  5385  5404CTCTACCTGCCAATCTCTGT  86 1239 1047943 N/A N/A  5497  5516GTTCCCCACGCCATTGTGTC  78 1240 1047959 N/A N/A  5788  5807CTCCTTCCCCCATTCTCTTG 104 1241 1047975 N/A N/A  5933  5952GCTACTACTAATAATAGCAA  99 1242 1047991 N/A N/A  6021  6040TTCGGCTCTCTCATCTGTCA  80 1243 1048007 N/A N/A  6117  6136ATGCAGGCTCCCAAGTGAGA  88 1244 1048023 N/A N/A  6280  6299CCACACTACATATAAGCTCT 163 1245 1048039 N/A N/A  6325  6344TCCTGCCTAGCCCAAATGCC 100 1246 1048055 N/A N/A  6403  6422TGTGCTTTTCTGCCTCCAGG  52 1247 1048071 N/A N/A  6543  6562TAGCCCTTTCTCCCCTGCCT  77 1248 1048087 N/A N/A  6958  6977ATCCAGAACCTTCCACACTG  88 1249 1048103 N/A N/A  7072  7091GGGACTTTTCCCAACAACTG  68 1250 1048119 N/A N/A  7393  7412CGTCCCTGGCCCTTCTCCCC  77 1251 1048135 N/A N/A  7843  7862CACAAGGCTGAAAAAGACTC  94 1252 1048151 N/A N/A  7987  8006GGAGGGTGACCCAAGTCCTT  35 1253 1048167 N/A N/A  8222  8241TGCCAGGAAAGTCTAACTCC  77 1254 1048183 N/A N/A  9362  9381TCCCCCCGCCCCGCCCGAGA  88 1255 1048199 N/A N/A  9533  9552CAGTATTACCTCTACTAGTC  64   20 1048215 N/A N/A  9609  9628CTGTCCCCTTTCCTCTTTCC  98 1256 1048231 N/A N/A  9788  9807CCAACCAGCCACATGACTCT  91 1257 1048247 N/A N/A  9825  9844TCAGGAGACCAGAGCTCAAG  93 1258 1048263 N/A N/A 10620 10639CCTGGCTTCATTTCAGCCCC  87 1259 1048279 N/A N/A 10711 10730GCAGAGAGAGCCTAGGCTCT 143 1260 1048295 N/A N/A 10792 10811GGCATGAGCCATCCTCTCCC 123 1261 1048311 N/A N/A 10941 10960ACTGGGCCCAAATCCCTCCT 104 1262 1048327 N/A N/A 11061 11080TTGCTCTCCTCCAGAATTCC  97 1263 1048343 N/A N/A 11226 11245CTACTAACTTTAATTCTCTT  97 1264 1048359 N/A N/A 11318 11337CTTCCCATTCCCCTGGTAGT  86 1265 1048375 N/A N/A 11487 11506GTCTTACTTTTCTTGATAGT  94 1266 1048391 N/A N/A 11610 11629TTGGACTCCTTAATGACCTG  86 1267

Example 2: Effect of 5-10-5 MOE Gapmer Modified Oligonucleotides onHuman GFAP RNA In Vitro, Single Dose

Modified oligonucleotides complementary to human GFAP nucleic acid weredesigned and tested for their single dose effects on GFAP RNA in vitro.The modified oligonucleotides were tested in a series of experimentsthat had similar culture conditions.

The modified oligonucleotides in the tables below are 5-10-5 MOE gapmerswith mixed PO/PS internucleoside linkages. The gapmers are 20nucleosides in length, wherein the central gap segment consists of ten2′-β-D-deoxynucleosides and the 5′ and 3′ wing segments each consists offive 2′-β-D-MOE modified nucleosides. The sugar motif for the gapmers is(from 5′ to 3′): eeeeeddddddddddeeeee; wherein ‘d’ represents a2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-β-D-MOE sugarmoiety. The internucleoside linkage motif for the gapmers is (from 5′ to3′): sooosssssssssssooss; wherein each ‘o’ represents a phosphodiesterinternucleoside linkage and each ‘s’ represents a phosphorothioateinternucleoside linkage. Each cytosine residue is a 5-methylcytosine.

“Start site” indicates the 5′-most nucleoside to which the modifiedoligonucleotide is complementary in the target nucleic acid sequence.“Stop site” indicates the 3′-most nucleoside to which the modifiedoligonucleotide is complementary in the target nucleic acid sequence.Each modified oligonucleotide listed in the Tables below is 100%complementary to SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 (GENBANKAccession No. NM_001131019.2). ‘N/A’ indicates that the modifiedoligonucleotide is not 100% complementary to that particular genesequence.

Cultured U251 cells were treated with modified oligonucleotide at aconcentration of 4,000 nM using free uptake at a density of 10,000 cellsper well. After a treatment period of approximately 48 hours, total RNAwas isolated from the cells and GFAP RNA levels were measured byquantitative real-time RTPCR. Human GFAP primer probe set RTS37485,described in Example 1 above, was used to measure RNA levels. GFAP RNAlevels were normalized to total RNA content, as measured by RIBOGREEN®.Results are presented in the tables below as percent GFAP RNA levelsrelative to untreated control cells. The values marked with an asterisk(*) indicate that the modified oligonucleotide is complementary to theamplicon region of the primer probe set. Additional assays may be usedto measure the potency and efficacy of the modified oligonucleotidescomplementary to the amplicon region.

TABLE 18Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleosidelinkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2NO: 2 Compound Start Stop Start Stop GFAP SEQ ID No. Site Site Site SiteSequence (5′ to 3′) (%UTC) NO 1047582 3047 3066 13336 13355GTCTTTATTTTTCCTCAGCG   8  517 1072810 1708 1727 11997 12016TCCATAACAACAGGAATCAG  65 1268 1072814 1719 1738 12008 12027TGGCAACAGTTTCCATAACA  31 1269 1072818 1728 1747 12017 12036CTCCATCTCTGGCAACAGTT  42 1270 1072822 1752 1771 12041 12060AGTTCCCAGATACTCCGAGA  56 1271 1072826 1763 1782 12052 12071CTCAAAGGCACAGTTCCCAG  56 1272 1072830 1786 1805 12075 12094TTTCCTCCAGCAGCCTGAGG  79 1273 1072834 1796 1815 12085 12104GAGTCTCAGTTTTCCTCCAG  18 1274 1072838 2173 2192 12462 12481GAATCAAGCTCCCACCTGCC  87 1275 1072842 2177 2196 12466 12485CTGAGAATCAAGCTCCCACC  94 1276 1072846 3039 3058 13328 13347TTTTCCTCAGCGACTAAAGG  48 1277 1072850 3059 3078 13348 13367GGCGCAGCATTTGTCTTTAT  45 1278 1072854 N/A N/A  8768  8787ATATCTTGTGACCTTGTGAT  53 1279 1072858 N/A N/A  8774  8793TTTGAGATATCTTGTGACCT  66 1280 1072862 N/A N/A  8780  8799GAGGCTTTTGAGATATCTTG  28 1281 1072866 N/A N/A  8785  8804ATTGTGAGGCTTTTGAGATA  66 1282 1072870 N/A N/A  7980  7999GACCCAAGTCCTTGGCCTTG  81 1283 1072874 N/A N/A  7990  8009TTTGGAGGGTGACCCAAGTC  78 1284 1072878 N/A N/A  7997  8016CTCTTAGTTTGGAGGGTGAC  61 1285 1072882 N/A N/A 11296 11315CCTTTTACCAAGCTGGAAAT  73 1286 1072886 N/A N/A 11303 11322GTAGTTTCCTTTTACCAAGC  34 1287 1072890 N/A N/A  4033  4052GAGACTTCTCGGGCACTCCT  75 1288 1072894 N/A N/A  4133  4152TGGGCCTGTTTCTGGTCCCT  63 1289 1072898 N/A N/A  4208  4227TTCCCCAGTAGGGAGGTGCT 100 1290 1072902 N/A N/A  4285  4304ATAGGTGAGCTCGCTGCCCA  82 1291 1072906 N/A N/A  4297  4316CACAGGCTCAGAATAGGTGA  68 1292 1072910 N/A N/A  4458  4477CAAGTCAAAGTAACTTGATG  80 1293 1072914 N/A N/A  4492  4511TGAATTTTATTATGACCACC  64 1294 1072918 N/A N/A  4564  4583CATGTCCTGTCAGCTCAGTG  61 1295 1072922 N/A N/A  4638  4657CACAAGCATACACTCACTGT  84 1296 1072926 N/A N/A  4677  4696CAGGGTTGGTGCACCTGCTT  74 1297 1072930 N/A N/A  4748  4767TAGACAGAGGACTTGTCTGG  96 1298 1072934 N/A N/A  4826  4845CCCTTGAGGCAGCTGTCACA  87 1299 1072938 N/A N/A  5049  5068CATTGCTCTGGCGGGCTGAG  89 1300 1072942 N/A N/A  5296  5315CAATCTCTGTGTTGAGCTTT  70 1301 1072946 N/A N/A  5396  5415TCATTTCCTGTCTCTACCTG  85 1302 1072950 N/A N/A  5549  5568GACCAGGGTGAGAGAAGCGG  78 1303 1072954 N/A N/A  5745  5764GAGGAGGCAGGCTGGCCCAC  84 1304 1072958 N/A N/A  5900  5919AATAATGGGTACTTTTGAAA  88 1305 1072962 N/A N/A  5986  6005TTCATAGTAAGGTAATCCAT  75 1306 1072966 N/A N/A  6032  6051CCTCTCTGGACTTCGGCTCT  81 1307 1072970 N/A N/A  6240  6259GGCACTATGTTTGGGTGCAC  85 1308 1072974 N/A N/A  6302  6321TCTACAGTGTCTTTCCTGGC  67 1309 1072978 N/A N/A  6446  6465CTAGGTGCCCTGGCTAGGCT  72 1310 1072982 N/A N/A  6524  6543TGCAGGGAGGTCCTCCCAGC  96 1311 1072986 N/A N/A  6901  6920GCGAGCGGAGGCCTGGGTGT  26 1312 1072990 N/A N/A  6942  6961ACTGACAGCTGCATCTGCGG  79 1313 1072994 N/A N/A  6985  7004AAGCGAATGAATGAACAGTG  69 1314 1072998 N/A N/A  7079  7098CCTGGCTGGGACTTTTCCCA  84 1315 1073002 N/A N/A  7119  7138GGGAGGTGAGCAGCACCCCA  84 1316 1073006 N/A N/A  7358  7377TGGCCGTCCCTGCTCCGCCC  96 1317 1073010 N/A N/A  7510  7529GGCCGGTCCCGCGGAGCCCC  86 1318 1073014 N/A N/A  7521  7540GGGATGGAGCCGGCCGGTCC  77 1319 1073018 N/A N/A  7785  7804AGCAGGGAGACTTCCCCAGG  85 1320 1073022 N/A N/A  7827  7846ACTCAGTCCCTGAAGGGAGC  90 1321 1073026 N/A N/A  7898  7917CTGCTATGTGTGAGGCAGGC  83 1322 1073030 N/A N/A  8027  8046CAATCTTGGCTGGGAAGATG  90 1323 1073034 N/A N/A  8048  8067AGATGGGTGAGGTGAGGAGT  33 1324 1073038 N/A N/A  8231  8250CCTTTTCCTTGCCAGGAAAG  76 1325 1073042 N/A N/A  9380  9399AGTAATTTAGCTCCCCCCTC  76 1326 1073046 N/A N/A  9410  9429AGAATCATTTCAGGGCCAAT  67 1327 1073050 N/A N/A  9438  9457GAAGAAGAGGAATTTTGTTC  82 1328 1073054 N/A N/A  9486  9505TTAAGTCCTGAGACATGCAT  55 1329 1073058 N/A N/A  9500  9519TTTCTGATGCTGAATTAAGT  88 1330 1073062 N/A N/A  9543  9562TAGGATTTGGCAGTATTACC  62 1331 1073066 N/A N/A  9649  9668TGTGGCACATATTAGTGCTC  76 1332 1073070 N/A N/A 10001 10020AATCCCCTTACTCGGGAGTC  76 1333 1073074 N/A N/A 10551 10570TTGAAATCAGGAGACCAGGA  73 1334 1073078 N/A N/A 10567 10586AAAACCCAGCACGGTATTGA  73 1335 1073082 N/A N/A 10715 10734CCGAGCAGAGAGAGCCTAGG  88 1336 1073086 N/A N/A 10805 10824CATGGACTTTCAGGGCATGA  90 1337 1073090 N/A N/A 11105 11124CAGCCTATGGAGGGACTGAG  88 1338 1073094 N/A N/A 11165 11184AAGAGAGAGTGTGTATTAGG  63 1339 1073098 N/A N/A 11208 11227TTTCTCTCCCTGGCAAGCAA  65 1340 1073102 N/A N/A 11256 11275AACTGTGTCTGCTAGAGTTG  66 1341 1073106 N/A N/A 11386 11405GTAAGCTGCTGGAGTAAGAT  45 1342 1073110 N/A N/A 11472 11491ATAGTAACCACAGCTGCCTT  81 1343 1073114 N/A N/A 11516 11535GTAACCTTGGGAAGTCCCCG  75 1344

TABLE 19 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG 7 517 1072811 1711 1730 12000 12019GTTTCCATAACAACAGGAAT 82 1345 1072815 1720 1739 12009 12028CTGGCAACAGTTTCCATAAC 40 1346 1072819 1746 1765 12035 12054CAGATACTCCGAGAGAACCT 75 1347 1072823 1753 1772 12042 12061CAGTTCCCAGATACTCCGAG 65 1348 1072827 1766 1785 12055 12074AAACTCAAAGGCACAGTTCC 99 1349 1072831 1787 1806 12076 12095TTTTCCTCCAGCAGCCTGAG 85 1350 1072835 1797 1816 12086 12105TGAGTCTCAGTTTTCCTCCA 18 1351 1072839 2174 2193 12463 12482AGAATCAAGCTCCCACCTGC 82 1352 1072843 3036 3055 13325 13344TCCTCAGCGACTAAAGGCAG 54 1353 1072847 3041 3060 13330 13349ATTTTTCCTCAGCGACTAAA 55 1354 1072851 3061 3080 13350 13369AGGGCGCAGCATTTGTCTTT 55 1355 1072855 N/A N/A 8771 8790GAGATATCTTGTGACCTTGT 37 1356 1072859 N/A N/A 8775 8794TTTTGAGATATCTTGTGACC 60 1357 1072863 N/A N/A 8781 8800TGAGGCTTTTGAGATATCTT 34 1358 1072867 N/A N/A 8786 8805TATTGTGAGGCTTTTGAGAT 60 1359 1072871 N/A N/A 7983 8002GGTGACCCAAGTCCTTGGCC 76 1360 1072875 N/A N/A 7991 8010GTTTGGAGGGTGACCCAAGT 70 1361 1072879 N/A N/A 11290 11309ACCAAGCTGGAAATGGAAAG 70 1362 1072883 N/A N/A 11297 11316TCCTTTTACCAAGCTGGAAA 83 1363 1072887 N/A N/A 11304 11323GGTAGTTTCCTTTTACCAAG 49 1364 1072891 N/A N/A 4035 4054CTGAGACTTCTCGGGCACTC 87 1365 1072895 N/A N/A 4162 4181GGCATGCGGGCATCAGATCC 87 1366 1072899 N/A N/A 4221 4240CTCCTGCACTGCTTTCCCCA 64 1367 1072903 N/A N/A 4287 4306GAATAGGTGAGCTCGCTGCC 89 1368 1072907 N/A N/A 4304 4323ATATGGACACAGGCTCAGAA 79 1369 1072911 N/A N/A 4463 4482CTGTGCAAGTCAAAGTAACT 76 1370 1072915 N/A N/A 4493 4512ATGAATTTTATTATGACCAC 67 1371 1072919 N/A N/A 4581 4600ACTTGAAGGCACACATGCAT 70 1372 1072923 N/A N/A 4648 4667CAGGCGCATCCACAAGCATA 88 1373 1072927 N/A N/A 4732 4751CTGGAGGATGAGCAGATGTG 57 1374 1072931 N/A N/A 4779 4798AGCAGCAGGAGGATTAAGGG 68 1375 1072935 N/A N/A 4827 4846TCCCTTGAGGCAGCTGTCAC 96 1376 1072939 N/A N/A 5058 5077GGAGCAGCACATTGCTCTGG 80 1377 1072943 N/A N/A 5299 5318TTGCAATCTCTGTGTTGAGC 55 1378 1072947 N/A N/A 5452 5471AGTTCGAATGCTCTCTTGTC 78 1379 1072951 N/A N/A 5715 5734ACCTTGGAGCGGTACCACTC 111 1380 1072955 N/A N/A 5828 5847TCAGTCACCTGGAGAGGATA 58 1381 1072959 N/A N/A 5918 5937AGCAATAGTAGCAGTAATAA 82 1382 1072963 N/A N/A 5987 6006CTTCATAGTAAGGTAATCCA 84 1383 1072967 N/A N/A 6122 6141ATGGAATGCAGGCTCCCAAG 71 1384 1072971 N/A N/A 6251 6270TGTTCTCTACGGGCACTATG 58 1385 1072975 N/A N/A 6374 6393CCACTAGGAATGGCCCTCCC 61 1386 1072979 N/A N/A 6451 6470CTCAGCTAGGTGCCCTGGCT 80 1387 1072983 N/A N/A 6561 6580CCTCACCCTGGGTTCTAATA 83 1388 1072987 N/A N/A 6903 6922AGGCGAGCGGAGGCCTGGGT 97 1389 1072991 N/A N/A 6944 6963ACACTGACAGCTGCATCTGC 79 1390 1072995 N/A N/A 6997 7016CACCTGGTCAGCAAGCGAAT 78 1391 1072999 N/A N/A 7082 7101GGCCCTGGCTGGGACTTTTC 79 1392 1073003 N/A N/A 7125 7144AAATCAGGGAGGTGAGCAGC 46 1393 1073007 N/A N/A 7382 7401CTTCTCCCCTGGCATCTCCT 77 1394 1073011 N/A N/A 7517 7536TGGAGCCGGCCGGTCCCGCG 87 1395 1073015 N/A N/A 7757 7776TGAGGGCTCACCGGTTCTCC 81 1396 1073019 N/A N/A 7789 7808AGGCAGCAGGGAGACTTCCC 88 1397 1073023 N/A N/A 7855 7874AAGGGATCTGCACACAAGGC 79 1398 1073027 N/A N/A 7924 7943TCAGTCATCAAACATCTAGT 78 1399 1073031 N/A N/A 8028 8047CCAATCTTGGCTGGGAAGAT 66 1400 1073035 N/A N/A 8051 8070AAGAGATGGGTGAGGTGAGG 36 1401 1073039 N/A N/A 8274 8293TAGGCTGGGTCTTGGTGCGG 76 1402 1073043 N/A N/A 9381 9400AAGTAATTTAGCTCCCCCCT 76 1403 1073047 N/A N/A 9411 9430AAGAATCATTTCAGGGCCAA 54 1404 1073051 N/A N/A 9440 9459CAGAAGAAGAGGAATTTTGT 70 1405 1073055 N/A N/A 9487 9506ATTAAGTCCTGAGACATGCA 64 1406 1073059 N/A N/A 9502 9521CCTTTCTGATGCTGAATTAA 72 1407 1073063 N/A N/A 9552 9571GTGACTATCTAGGATTTGGC 23 1408 1073067 N/A N/A 9685 9704GAGGAGACAATTAACTAAAA 64 1409 1073071 N/A N/A 10125 10144TCGAAAGCAGGCAAGCAAGC 99 1410 1073075 N/A N/A 10552 10571ATTGAAATCAGGAGACCAGG 66 1411 1073079 N/A N/A 10612 10631CATTTCAGCCCCTCTGCAAG 77 1412 1073083 N/A N/A 10724 10743CCTATGCAACCGAGCAGAGA 95 1413 1073087 N/A N/A 10848 10867GTGTCCAGGCTGGTTTCTGC 82 1414 1073091 N/A N/A 11124 11143ACCAAGGCTCCCCTTAGAAC 59 1415 1073095 N/A N/A 11166 11185AAAGAGAGAGTGTGTATTAG 78 1416 1073099 N/A N/A 11210 11229TCTTTCTCTCCCTGGCAAGC 73 1417 1073103 N/A N/A 11283 11302TGGAAATGGAAAGCCCTCCC 78 1418 1073107 N/A N/A 11391 11410GAGTGGTAAGCTGCTGGAGT 42 1419 1073111 N/A N/A 11474 11493TGATAGTAACCACAGCTGCC 92 1420 1073115 N/A N/A 11518 11537GTGTAACCTTGGGAAGTCCC 77 1421

TABLE 20 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG 7 517 1072812 1713 1732 12002 12021CAGTTTCCATAACAACAGGA 58 1422 1072816 1722 1741 12011 12030CTCTGGCAACAGTTTCCATA 69 1423 1072820 1749 1768 12038 12057TCCCAGATACTCCGAGAGAA 73 1424 1072824 1758 1777 12047 12066AGGCACAGTTCCCAGATACT 36 1425 1072828 1784 1803 12073 12092TCCTCCAGCAGCCTGAGGAA 71 1426 1072832 1788 1807 12077 12096GTTTTCCTCCAGCAGCCTGA 64 1427 1072836 2168 2187 12457 12476AAGCTCCCACCTGCCCACAG 73 1428 1072840 2175 2194 12464 12483GAGAATCAAGCTCCCACCTG 74 1429 1072844 3037 3056 13326 13345TTCCTCAGCGACTAAAGGCA 61 1430 1072848 3042 3061 13331 13350TATTTTTCCTCAGCGACTAA 69 1431 1072852 3063 3082 13352 13371GAAGGGCGCAGCATTTGTCT 57 1432 1072856 N/A N/A 8772 8791TGAGATATCTTGTGACCTTG 43 1433 1072860 N/A N/A 8776 8795CTTTTGAGATATCTTGTGAC 62 1434 1072864 N/A N/A 8783 8802TGTGAGGCTTTTGAGATATC 43 1435 1072868 N/A N/A 8788 8807CGTATTGTGAGGCTTTTGAG 20 1436 1072872 N/A N/A 7984 8003GGGTGACCCAAGTCCTTGGC 14 1437 1072876 N/A N/A 7992 8011AGTTTGGAGGGTGACCCAAG 61 1438 1072880 N/A N/A 11293 11312TTTACCAAGCTGGAAATGGA 83 1439 1072884 N/A N/A 11298 11317TTCCTTTTACCAAGCTGGAA 77 1440 1072888 N/A N/A 11310 11329TCCCCTGGTAGTTTCCTTTT 85 1441 1072892 N/A N/A 4055 4074CAGGGAGGTTCGGCCCCTCC 83 1442 1072896 N/A N/A 4174 4193CTCCTGGCAGAAGGCATGCG 75 1443 1072900 N/A N/A 4231 4250GGCCCCGCTGCTCCTGCACT 105 1444 1072904 N/A N/A 4289 4308CAGAATAGGTGAGCTCGCTG 83 1445 1072908 N/A N/A 4307 4326TTTATATGGACACAGGCTCA 83 1446 1072912 N/A N/A 4482 4501TATGACCACCGCTTCACAGC 95 1447 1072916 N/A N/A 4558 4577CTGTCAGCTCAGTGAAGCGC 86 1448 1072920 N/A N/A 4628 4647CACTCACTGTTGCACACACA 89 1449 1072924 N/A N/A 4651 4670ACACAGGCGCATCCACAAGC 98 1450 1072928 N/A N/A 4739 4758GACTTGTCTGGAGGATGAGC 73 1451 1072932 N/A N/A 4785 4804CAGTAGAGCAGCAGGAGGAT 82 1452 1072936 N/A N/A 4882 4901GGACACATTCCTGGGTCCAG 100 1453 1072940 N/A N/A 5142 5161GGTGAGGAGTAGAGGGCCAC 88 1454 1072944 N/A N/A 5307 5326TCTCTCAGTTGCAATCTCTG 84 1455 1072948 N/A N/A 5453 5472GAGTTCGAATGCTCTCTTGT 75 1456 1072952 N/A N/A 5730 5749CCCACAGGCAGGGCTACCTT 89 1457 1072956 N/A N/A 5891 5910TACTTTTGAAAGCAATAGTG 77 1458 1072960 N/A N/A 5953 5972CTTAGAACAGAACAGTATCA 101 1459 1072964 N/A N/A 6028 6047TCTGGACTTCGGCTCTCTCA 55 1460 1072968 N/A N/A 6140 6159AAACAGACTGGCAGAGGCAT 59 1461 1072972 N/A N/A 6260 6279GAGCTGTGGTGTTCTCTACG 58 1462 1072976 N/A N/A 6380 6399TGTCCTCCACTAGGAATGGC 76 1463 1072980 N/A N/A 6459 6478TCACACTCCTCAGCTAGGTG 32 1464 1072984 N/A N/A 6563 6582GGCCTCACCCTGGGTTCTAA 76 1465 1072988 N/A N/A 6905 6924TTAGGCGAGCGGAGGCCTGG 95 1466 1072992 N/A N/A 6945 6964CACACTGACAGCTGCATCTG 90 1467 1072996 N/A N/A 7003 7022GCACAACACCTGGTCAGCAA 73 1468 1073000 N/A N/A 7085 7104GTTGGCCCTGGCTGGGACTT 63 1469 1073004 N/A N/A 7126 7145GAAATCAGGGAGGTGAGCAG 63 1470 1073008 N/A N/A 7475 7494GGTTTCGAGGCCCGGCCCCC 73 1471 1073012 N/A N/A 7518 7537ATGGAGCCGGCCGGTCCCGC 92 1472 1073016 N/A N/A 7762 7781TGTGATGAGGGCTCACCGGT 65 1473 1073020 N/A N/A 7797 7816CTACCGTGAGGCAGCAGGGA 86 1474 1073024 N/A N/A 7876 7895TGTGCTGGGCATTGAGGTGG 64 1475 1073028 N/A N/A 7967 7986GGCCTTGAGGCCTAATCAAT 78 1476 1073032 N/A N/A 8045 8064TGGGTGAGGTGAGGAGTCCA 73 1477 1073036 N/A N/A 8057 8076AGGCAGAAGAGATGGGTGAG 67 1478 1073040 N/A N/A 9355 9374GCCCCGCCCGAGAGAGAAAA 92 1479 1073044 N/A N/A 9407 9426ATCATTTCAGGGCCAATGCA 66 1480 1073048 N/A N/A 9423 9442TGTTCCTTAGCTAAGAATCA 58 1481 1073052 N/A N/A 9442 9461TCCAGAAGAAGAGGAATTTT 94 1482 1073056 N/A N/A 9488 9507AATTAAGTCCTGAGACATGC 51 1483 1073060 N/A N/A 9517 9536AGTCAGCCTGGTTAGCCTTT 36 1484 1073064 N/A N/A 9560 9579AGTGACCTGTGACTATCTAG 35 1485 1073068 N/A N/A 9889 9908TCTGCCGAAGGAAGGAAGGA 77 1486 1073072 N/A N/A 10129 10148TCCGTCGAAAGCAGGCAAGC 83 1487 1073076 N/A N/A 10555 10574GGTATTGAAATCAGGAGACC 59 1488 1073080 N/A N/A 10662 10681CAATAGTGCTGCTGCCAGAG 54 1489 1073084 N/A N/A 10727 10746GAACCTATGCAACCGAGCAG 56 1490 1073088 N/A N/A 11008 11027TATGTGCCAGCCCCAGGCTT 89 1491 1073092 N/A N/A 11128 11147TGGTACCAAGGCTCCCCTTA 74 1492 1073096 N/A N/A 11187 11206GCCCCCGAGTTTGAGGGTGA 90 1493 1073100 N/A N/A 11240 11259GTTGGAAGTGAAAGCTACTA 58 1494 1073104 N/A N/A 11365 11384AGCTCCCACTGTGGTTGGAG 87 1495 1073108 N/A N/A 11399 11418TGCCTGGCGAGTGGTAAGCT 91 1496 1073112 N/A N/A 11476 11495CTTGATAGTAACCACAGCTG 63 1497 1073116 N/A N/A 11561 11580AGTTAGGAGTTCACACAGAC 72 1498

TABLE 21 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG 8 517 1072813 1717 1736 12006 12025GCAACAGTTTCCATAACAAC 32 1499 1072817 1725 1744 12014 12033CATCTCTGGCAACAGTTTCC 62 1500 1072821 1751 1770 12040 12059GTTCCCAGATACTCCGAGAG 53 1501 1072825 1759 1778 12048 12067AAGGCACAGTTCCCAGATAC 51 1502 1072829 1785 1804 12074 12093TTCCTCCAGCAGCCTGAGGA 89 1503 1072833 1789 1808 12078 12097AGTTTTCCTCCAGCAGCCTG 46 1504 1072837 2171 2190 12460 12479ATCAAGCTCCCACCTGCCCA 64 1505 1072841 2176 2195 12465 12484TGAGAATCAAGCTCCCACCT 90 1506 1072845 3038 3057 13327 13346TTTCCTCAGCGACTAAAGGC 52 1507 1072849 3057 3076 13346 13365CGCAGCATTTGTCTTTATTT 26 1508 1072853 3064 3083 13353 13372GGAAGGGCGCAGCATTTGTC 43 1509 1072857 N/A N/A 8773 8792TTGAGATATCTTGTGACCTT 32 1510 1072861 N/A N/A 8779 8798AGGCTTTTGAGATATCTTGT 35 1511 1072865 N/A N/A 8784 8803TTGTGAGGCTTTTGAGATAT 47 1512 1072869 N/A N/A 7977 7996CCAAGTCCTTGGCCTTGAGG 93 1513 1072873 N/A N/A 7988 8007TGGAGGGTGACCCAAGTCCT 107 1514 1072877 N/A N/A 7994 8013TTAGTTTGGAGGGTGACCCA 68 1515 1072881 N/A N/A 11295 11314CTTTTACCAAGCTGGAAATG 103 1516 1072885 N/A N/A 11299 11318TTTCCTTTTACCAAGCTGGA 80 1517 1072889 N/A N/A 4027 4046TCTCGGGCACTCCTTCTTGG 96 1518 1072893 N/A N/A 4084 4103CCCATCAAGAGGTAGGGAGG 66 1519 1072897 N/A N/A 4183 4202GACCCTGGACTCCTGGCAGA 83 1520 1072901 N/A N/A 4253 4272CCAGCCATGAATGAAACACA 82 1521 1072905 N/A N/A 4293 4312GGCTCAGAATAGGTGAGCTC 67 1522 1072909 N/A N/A 4435 4454GTCACAAGCTGGTGGCAGGC 69 1523 1072913 N/A N/A 4487 4506TTTATTATGACCACCGCTTC 92 1524 1072917 N/A N/A 4563 4582ATGTCCTGTCAGCTCAGTGA 50 1525 1072921 N/A N/A 4632 4651CATACACTCACTGTTGCACA 83 1526 1072925 N/A N/A 4669 4688GTGCACCTGCTTCTGCTCAC 111 1527 1072929 N/A N/A 4741 4760AGGACTTGTCTGGAGGATGA 47 1528 1072933 N/A N/A 4822 4841TGAGGCAGCTGTCACAGAGA 102 1529 1072937 N/A N/A 4904 4923CATCCTGGAGCCTGGAGTGG 87 1530 1072941 N/A N/A 5295 5314AATCTCTGTGTTGAGCTTTC 56 1531 1072945 N/A N/A 5308 5327CTCTCTCAGTTGCAATCTCT 75 1532 1072949 N/A N/A 5460 5479CGGCCAGGAGTTCGAATGCT 81 1533 1072953 N/A N/A 5733 5752TGGCCCACAGGCAGGGCTAC 83 1534 1072957 N/A N/A 5895 5914TGGGTACTTTTGAAAGCAAT 75 1535 1072961 N/A N/A 5965 5984AAAAGCACAGGGCTTAGAAC 76 1536 1072965 N/A N/A 6031 6050CTCTCTGGACTTCGGCTCTC 88 1537 1072969 N/A N/A 6167 6186GGCATATGGTAGAGGCTCAG 56 1538 1072973 N/A N/A 6270 6289TATAAGCTCTGAGCTGTGGT 37 1539 1072977 N/A N/A 6443 6462GGTGCCCTGGCTAGGCTAGC 77 1540 1072981 N/A N/A 6460 6479CTCACACTCCTCAGCTAGGT 77 1541 1072985 N/A N/A 6897 6916GCGGAGGCCTGGGTGTTTTG 75 1542 1072989 N/A N/A 6908 6927GTCTTAGGCGAGCGGAGGCC 90 1543 1072993 N/A N/A 6972 6991AACAGTGCCACAGAATCCAG 91 1544 1072997 N/A N/A 7051 7070GACCCATGGATGCGGGCAGG 68 1545 1073001 N/A N/A 7086 7105CGTTGGCCCTGGCTGGGACT 71 1546 1073005 N/A N/A 7201 7220TCTGTCAGGTCTGCAAACTA 83 1547 1073009 N/A N/A 7479 7498GGGAGGTTTCGAGGCCCGGC 41 1548 1073013 N/A N/A 7520 7539GGATGGAGCCGGCCGGTCCC 81 1549 1073017 N/A N/A 7764 7783GCTGTGATGAGGGCTCACCG 68 1550 1073021 N/A N/A 7818 7837CTGAAGGGAGCAAGATGAGC 82 1551 1073025 N/A N/A 7878 7897ACTGTGCTGGGCATTGAGGT 77 1552 1073029 N/A N/A 8005 8024GAGTATGCCTCTTAGTTTGG 63 1553 1073033 N/A N/A 8046 8065ATGGGTGAGGTGAGGAGTCC 36 1554 1073037 N/A N/A 8078 8097GTGGTGAAGAAAGTTCCAAG 88 1555 1073041 N/A N/A 9356 9375CGCCCCGCCCGAGAGAGAAA 64 1556 1073045 N/A N/A 9409 9428GAATCATTTCAGGGCCAATG 27 1557 1073049 N/A N/A 9425 9444TTTGTTCCTTAGCTAAGAAT 62 1558 1073053 N/A N/A 9444 9463AGTCCAGAAGAAGAGGAATT 68 1559 1073057 N/A N/A 9497 9516CTGATGCTGAATTAAGTCCT 50 1560 1073061 N/A N/A 9519 9538CTAGTCAGCCTGGTTAGCCT 57 1561 1073065 N/A N/A 9577 9596CCATTTATCTGTGCTTTAGT 39 1562 1073069 N/A N/A 9892 9911CGCTCTGCCGAAGGAAGGAA 61 1563 1073073 N/A N/A 10548 10567AAATCAGGAGACCAGGAGGG 59 1564 1073077 N/A N/A 10561 10580CAGCACGGTATTGAAATCAG 42 1565 1073081 N/A N/A 10692 10711TTCCAAACGGGCTGGAGAGC 84 1566 1073085 N/A N/A 10804 10823ATGGACTTTCAGGGCATGAG 64 1567 1073089 N/A N/A 11097 11116GGAGGGACTGAGGAAACGGA 68 1568 1073093 N/A N/A 11157 11176GTGTGTATTAGGATCCCATC 24 1569 1073097 N/A N/A 11193 11212AGCAAGGCCCCCGAGTTTGA 81 1570 1073101 N/A N/A 11246 11265GCTAGAGTTGGAAGTGAAAG 67 1571 1073105 N/A N/A 11368 11387ATGAGCTCCCACTGTGGTTG 72 1572 1073109 N/A N/A 11427 11446GATAACACTGGGAAAGCATT 91 1573 1073113 N/A N/A 11514 11533AACCTTGGGAAGTCCCCGAC 85 1574 1073117 N/A N/A 11563 11582ACAGTTAGGAGTTCACACAG 75 1575

TABLE 22 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixed PO/PSinternucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ ID NO: 1NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No. SiteSite Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 3066 1333613355 GTCTTTATTTTTCCTCAGCG  5 517 1103152 41 60 3489 3508CTGGCTCTGCTCGCTCCTGG 91 1576 1103168 272 291 3720 3739CTCCATCATCTCTGCCCGCT 106  1577 1103184 565 584 4950 4969CTATAGGCAGCCAGGTTGTT  69* 1578 1103200 760 779 5646 5665ATCTCTTTCAGGGCTGCGGT 65 1579 1103216 1062 1081 7634 7653TGAGGCTCTGCCCCTCTTCC 111  1580 1103232 1335 1354 11624 11643CCTTGTGCTCCTGCTTGGAC 88 1581 1103248 1527 1546 11816 11835AGGCCTGATACTGACGGAGC 79 1582 1103264 1669 1688 11958 11977GTAGGTGCCCCCCGCCCTCC 90 1583 1103280 1733 1752 12022 12041AGAACCTCCATCTCTGGCAA 62 1584 1103296 1860 1879 12149 12168CCAAAAGACAAAACAAGCCT 75 1585 1103312 1886 1905 12175 12194CATAGGGATATCCCACCTCA 111  1586 1103328 2094 2113 12383 12402GTCATCGCTCAGGAGGTCCT 69 1587 1103344 2231 2250 12520 12539CTATCCCTCCCAGCACCTCA 109  1588 1103360 2427 2446 12716 12735AGGATGAGTCACTTCCTTAA 73 1589 1103376 2484 2503 12773 12792GTCATGCCCTGCCCCCATGG 90 1590 1103392 2512 2531 12801 12820AGGAAGAGGCCTTTAGAAAT 68 1591 1103408 2672 2691 12961 12980GTGTGTGAGTAAGAAGGGAC 55 1592 1103424 2724 2743 13013 13032TCAGTTTTACAATTGTAAAA 83 1593 1103440 2899 2918 13188 13207GGAGAACCCTGAAGTGGGCC 76 1594 1103472 N/A N/A 8787 8806GTATTGTGAGGCTTTTGAGA 43 1595 1103488 N/A N/A 8899 8918GAGGCTCACTCCCTGTCAAG 60 1596 1103504 N/A N/A 9045 9064AACAAGCTCTGCCAGTTTAA 66 1597 1103520 N/A N/A 9234 9253TGAGTCAGCACTGAGCTGAG 95 1598 1103536 N/A N/A 9270 9289CAAGAGCTGCGGTCCTGAGG 59 1599 1103552 N/A N/A 9310 9329TTTAACATTAAGAGCAGGGA 63 1600 1103568 N/A N/A 8532 8551CTGGTATGATAGGCTCTGGC 86 1601 1103584 N/A N/A 8650 8669CAGACAGGGCAGATGTCAAG 92 1602 1103600 N/A N/A 3999 4018CCCCTTCTGCTCACAAGGCC 121  1603 1103616 N/A N/A 4160 4179CATGCGGGCATCAGATCCCC 89 1604 1103632 N/A N/A 4275 4294TCGCTGCCCACAGTCACAAA 104  1605 1103648 N/A N/A 4407 4426GTGCTTTGCGCCCAGACCTG 105  1606 1103664 N/A N/A 4496 4515GAAATGAATTTTATTATGAC 106  1607 1103680 N/A N/A 4640 4659TCCACAAGCATACACTCACT 81 1608 1103696 N/A N/A 4837 4856TGCCTCAGTCTCCCTTGAGG 107  1609 1103712 N/A N/A 5100 5119CCATTCCTCAGCCTTGCCTT 98 1610 1103728 N/A N/A 5346 5365GTCTCTCTCAGTCTCAGCTT 78 1611 1103744 N/A N/A 5413 5432GTCTTTCTGTTTGTCTTTCA 57 1612 1103760 N/A N/A 5487 5506CCATTGTGTCCTCTTCTGCC 102  1613 1103776 N/A N/A 5776 5795TTCTCTTGTACAGAGCAAGA 79 1614 1103792 N/A N/A 5860 5879CTGGGCAAGCCATCTCACTT 86 1615 1103808 N/A N/A 5941 5960CAGTATCAGCTACTACTAAT 80 1616 1103824 N/A N/A 5978 5997AAGGTAATCCATGAAAAGCA 84 1617 1103840 N/A N/A 6024 6043GACTTCGGCTCTCTCATCTG 82 1618 1103856 N/A N/A 6204 6223CCAGGTCAGACACCTCTCTG 81 1619 1103872 N/A N/A 6267 6286AAGCTCTGAGCTGTGGTGTT 33 1620 1103888 N/A N/A 6315 6334CCCAAATGCCCCCTCTACAG 93 1621 1103904 N/A N/A 6414 6433CCCTGCCTCTCTGTGCTTTT 92 1622 1103920 N/A N/A 6517 6536AGGTCCTCCCAGCCCCATCG 108  1623 1103936 N/A N/A 6947 6966TCCACACTGACAGCTGCATC 92 1624 1103952 N/A N/A 7030 7049TAGCGGGCTGCCAGACCTCA 102  1625 1103968 N/A N/A 7326 7345CCCTGGCCGCGCTCACCGTG 96 1626 1103984 N/A N/A 7421 7440TTCAGGCCCCGCCCTCGACC 81 1627 1104000 N/A N/A 7814 7833AGGGAGCAAGATGAGCTCTA 120  1628 1104016 N/A N/A 7940 7959GAATCCATCCATCCATTCAG 86 1629 1104032 N/A N/A 8038 8057GGTGAGGAGTCCAATCTTGG 76 1630 1104048 N/A N/A 8080 8099AAGTGGTGAAGAAAGTTCCA 108  1631 1104064 N/A N/A 8251 8270CATCATGACAACTTGAACGC 98 1632 1104080 N/A N/A 9364 9383CCTCCCCCCGCCCCGCCCGA 111  1633 1104096 N/A N/A 9414 9433GCTAAGAATCATTTCAGGGC 40 1634 1104112 N/A N/A 9455 9474CTAAATATTCTAGTCCAGAA 97 1635 1104128 N/A N/A 9509 9528TGGTTAGCCTTTCTGATGCT 84 1636 1104144 N/A N/A 9535 9554GGCAGTATTACCTCTACTAG 28 1637 1104160 N/A N/A 9559 9578GTGACCTGTGACTATCTAGG 47 1638 1104176 N/A N/A 9579 9598TGCCATTTATCTGTGCTTTA 50 1639 1104192 N/A N/A 9665 9684TAAAGGCTGTTAAACATGTG 51 1640 1104208 N/A N/A 9785 9804ACCAGCCACATGACTCTGGG 83 1641 1104224 N/A N/A 10558 10577CACGGTATTGAAATCAGGAG 47 1642 1104240 N/A N/A 10654 10673CTGCTGCCAGAGTCCTGGCT 88 1643 1104256 N/A N/A 10753 10772CACCCCCCTCCCCATCATGA 89 1644 1104272 N/A N/A 10818 10837AGAGGAGGCCTCTCATGGAC 96 1645 1104288 N/A N/A 11083 11102AACGGAATTACATTCAGTTT 81 1646 1104304 N/A N/A 11152 11171TATTAGGATCCCATCTAGTG 103  1647 1104320 N/A N/A 11230 11249AAAGCTACTAACTTTAATTC 81 1648 1104336 N/A N/A 11288 11307CAAGCTGGAAATGGAAAGCC 83 1649 1104352 N/A N/A 11390 11409AGTGGTAAGCTGCTGGAGTA 85 1650 1104368 N/A N/A 11499 11518CCGACTTCCCAGGTCTTACT 78 1651

TABLE 23 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  6 517 1103150 20 39 3468 3487ATGCGAGGGCTTTATGAAGG 82 1652 1103166 264 283 3712 3731TCTCTGCCCGCTCACTGGCC 126  1653 1103182 549 568 4934 4953TGTTCTCGGCTTCCAGCCTC  40* 1654 1103198 758 777 5644 5663CTCTTTCAGGGCTGCGGTGA 35 1655 1103214 1049 1068 7621 7640CTCTTCCTCCAGCCGCGCCA 70 1656 1103230 1327 1346 11616 11635TCCTGCTTGGACTCCTTAAT 69 1657 1103246 1501 1520 11790 11809CAGCAAGCTGACCTAGGGAC 39 1658 1103262 1662 1681 11951 11970CCCCCCGCCCTCCTCCCCTT 91 1659 1103278 1729 1748 12018 12037CCTCCATCTCTGGCAACAGT 34 1660 1103294 1858 1877 12147 12166AAAAGACAAAACAAGCCTCT 81 1661 1103310 1884 1903 12173 12192TAGGGATATCCCACCTCATA 74 1662 1103326 2061 2080 12350 12369CAGGTGACTGCCCCAGGTGG 73 1663 1103342 2228 2247 12517 12536TCCCTCCCAGCACCTCATCC 80 1664 1103358 2424 2443 12713 12732ATGAGTCACTTCCTTAATTC 58 1665 1103374 2467 2486 12756 12775TGGATACATCCCCTTTCTCT 59 1666 1103390 2510 2529 12799 12818GAAGAGGCCTTTAGAAATGG 76 1667 1103406 2668 2687 12957 12976GTGAGTAAGAAGGGACCGCA 82 1668 1103422 2717 2736 13006 13025TACAATTGTAAAATAGGGCA 90 1669 1103438 2849 2868 13138 13157ACCAGTCTGCTCACCAGTCT 74 1670 1103470 N/A N/A 8769 8788GATATCTTGTGACCTTGTGA 34 1671 1103486 N/A N/A 8897 8916GGCTCACTCCCTGTCAAGCT 100  1672 1103502 N/A N/A 9043 9062CAAGCTCTGCCAGTTTAATG 30 1673 1103518 N/A N/A 9224 9243CTGAGCTGAGCGATGGAGCC 71 1674 1103534 N/A N/A 9263 9282TGCGGTCCTGAGGGAAGAAT 76 1675 1103550 N/A N/A 9308 9327TAACATTAAGAGCAGGGAAC 77 1676 1103566 N/A N/A 8500 8519AGTGTCACGAAGGCCCCCAG 49 1677 1103582 N/A N/A 8584 8603GGACCAGGGCCTAGCAGGAC 69 1678 1103598 N/A N/A 3993 4012CTGCTCACAAGGCCCCCCTT 92 1679 1103614 N/A N/A 4049 4068GGTTCGGCCCCTCCCTGAGA 69 1680 1103630 N/A N/A 4250 4269GCCATGAATGAAACACAGGG 66 1681 1103646 N/A N/A 4391 4410CCTGCCTGCTCTTTCCCTCA 76 1682 1103662 N/A N/A 4490 4509AATTTTATTATGACCACCGC 110  1683 1103678 N/A N/A 4635 4654AAGCATACACTCACTGTTGC 78 1684 1103694 N/A N/A 4817 4836CAGCTGTCACAGAGACCACC 100  1685 1103710 N/A N/A 5097 5116TTCCTCAGCCTTGCCTTACC 78 1686 1103726 N/A N/A 5305 5324TCTCAGTTGCAATCTCTGTG 87 1687 1103742 N/A N/A 5398 5417TTTCATTTCCTGTCTCTACC 89 1688 1103758 N/A N/A 5481 5500TGTCCTCTTCTGCCTGCCCC 94 1689 1103774 N/A N/A 5726 5745CAGGCAGGGCTACCTTGGAG 87 1690 1103790 N/A N/A 5849 5868ATCTCACTTCTCTGGTGAAA 95 1691 1103806 N/A N/A 5923 5942ATAATAGCAATAGTAGCAGT 103  1692 1103822 N/A N/A 5974 5993TAATCCATGAAAAGCACAGG 84 1693 1103838 N/A N/A 6020 6039TCGGCTCTCTCATCTGTCAA 53 1694 1103854 N/A N/A 6199 6218TCAGACACCTCTCTGTGTCC 86 1695 1103870 N/A N/A 6257 6276CTGTGGTGTTCTCTACGGGC 78 1696 1103886 N/A N/A 6313 6332CAAATGCCCCCTCTACAGTG 105  1697 1103902 N/A N/A 6408 6427CTCTCTGTGCTTTTCTGCCT 71 1698 1103918 N/A N/A 6501 6520ATCGGGCCCTCACCCTGCTC 107  1699 1103934 N/A N/A 6904 6923TAGGCGAGCGGAGGCCTGGG 90 1700 1103950 N/A N/A 7017 7036GACCTCAGCACCTAGCACAA 82 1701 1103966 N/A N/A 7314 7333TCACCGTGCCGCGCAGAGAC 101  1702 1103982 N/A N/A 7416 7435GCCCCGCCCTCGACCCAGGT 88 1703 1103998 N/A N/A 7805 7824GATGAGCTCTACCGTGAGGC 71 1704 1104014 N/A N/A 7922 7941AGTCATCAAACATCTAGTGA 46 1705 1104030 N/A N/A 8035 8054GAGGAGTCCAATCTTGGCTG 67 1706 1104046 N/A N/A 8075 8094GTGAAGAAAGTTCCAAGGAG 93 1707 1104062 N/A N/A 8228 8247TTTCCTTGCCAGGAAAGTCT 81 1708 1104078 N/A N/A 9359 9378CCCCGCCCCGCCCGAGAGAG 36 1709 1104094 N/A N/A 9412 9431TAAGAATCATTTCAGGGCCA 71 1710 1104110 N/A N/A 9452 9471AATATTCTAGTCCAGAAGAA 103  1711 1104126 N/A N/A 9506 9525TTAGCCTTTCTGATGCTGAA 58 1712 1104142 N/A N/A 9530 9549TATTACCTCTACTAGTCAGC 65 1713 1104158 N/A N/A 9557 9576GACCTGTGACTATCTAGGAT 18 1714 1104174 N/A N/A 9576 9595CATTTATCTGTGCTTTAGTG 37 1715 1104190 N/A N/A 9660 9679GCTGTTAAACATGTGGCACA 74 1716 1104206 N/A N/A 9706 9725CCTACTTCTCTAGGTGGGAG 65 1717 1104222 N/A N/A 10556 10575CGGTATTGAAATCAGGAGAC 47 1718 1104238 N/A N/A 10610 10629TTTCAGCCCCTCTGCAAGCC 68 1719 1104254 N/A N/A 10747 10766CCTCCCCATCATGAGTATGA 92 1720 1104270 N/A N/A 10816 10835AGGAGGCCTCTCATGGACTT 93 1721 1104286 N/A N/A 11078 11097AATTACATTCAGTTTCCTTG 88 1722 1104302 N/A N/A 11150 11169TTAGGATCCCATCTAGTGGC 60 1723 1104318 N/A N/A 11225 11244TACTAACTTTAATTCTCTTT 49 1724 1104334 N/A N/A 11280 11299AAATGGAAAGCCCTCCCCAT 98 1725 1104350 N/A N/A 11388 11407TGGTAAGCTGCTGGAGTAAG 55 1726 1104366 N/A N/A 11494 11513TTCCCAGGTCTTACTTTTCT 76 1727

TABLE 24 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  6 517 1103151 39 58 3487 3506GGCTCTGCTCGCTCCTGGGA 108  1728 1103167 269 288 3717 3736CATCATCTCTGCCCGCTCAC 93 1729 1103183 553 572 4938 4957AGGTTGTTCTCGGCTTCCAG  55* 1730 1103199 759 778 5645 5664TCTCTTTCAGGGCTGCGGTG 54 1731 1103215 1050 1069 7622 7641CCTCTTCCTCCAGCCGCGCC 64 1732 1103231 1334 1353 11623 11642CTTGTGCTCCTGCTTGGACT 53 1733 1103247 1518 1537 11807 11826ACTGACGGAGCCTAGGGCAG 61 1734 1103263 1667 1686 11956 11975AGGTGCCCCCCGCCCTCCTC 98 1735 1103279 1730 1749 12019 12038ACCTCCATCTCTGGCAACAG 25 1736 1103295 1859 1878 12148 12167CAAAAGACAAAACAAGCCTC 73 1737 1103311 1885 1904 12174 12193ATAGGGATATCCCACCTCAT 75 1738 1103327 2086 2105 12375 12394TCAGGAGGTCCTTCTGGGAT 67 1739 1103343 2230 2249 12519 12538TATCCCTCCCAGCACCTCAT 63 1740 1103359 2425 2444 12714 12733GATGAGTCACTTCCTTAATT 35 1741 1103375 2483 2502 12772 12791TCATGCCCTGCCCCCATGGA 69 1742 1103391 2511 2530 12800 12819GGAAGAGGCCTTTAGAAATG 74 1743 1103407 2670 2689 12959 12978GTGTGAGTAAGAAGGGACCG 46 1744 1103423 2722 2741 13011 13030AGTTTTACAATTGTAAAATA 88 1745 1103439 2868 2887 13157 13176ATCTCTGGGCACAGATCCCA 93 1746 1103471 N/A N/A 8770 8789AGATATCTTGTGACCTTGTG 35 1747 1103487 N/A N/A 8898 8917AGGCTCACTCCCTGTCAAGC 62 1748 1103503 N/A N/A 9044 9063ACAAGCTCTGCCAGTTTAAT 51 1749 1103519 N/A N/A 9225 9244ACTGAGCTGAGCGATGGAGC 65 1750 1103535 N/A N/A 9266 9285AGCTGCGGTCCTGAGGGAAG 59 1751 1103551 N/A N/A 9309 9328TTAACATTAAGAGCAGGGAA 68 1752 1103567 N/A N/A 8530 8549GGTATGATAGGCTCTGGCTA 21 1753 1103583 N/A N/A 8649 8668AGACAGGGCAGATGTCAAGC 90 1754 1103599 N/A N/A 3994 4013TCTGCTCACAAGGCCCCCCT 56 1755 1103615 N/A N/A 4053 4072GGGAGGTTCGGCCCCTCCCT 94 1756 1103631 N/A N/A 4251 4270AGCCATGAATGAAACACAGG 115  1757 1103647 N/A N/A 4401 4420TGCGCCCAGACCTGCCTGCT 75 1758 1103663 N/A N/A 4495 4514AAATGAATTTTATTATGACC 80 1759 1103679 N/A N/A 4636 4655CAAGCATACACTCACTGTTG 69 1760 1103695 N/A N/A 4820 4839AGGCAGCTGTCACAGAGACC 95 1761 1103711 N/A N/A 5099 5118CATTCCTCAGCCTTGCCTTA 95 1762 1103727 N/A N/A 5311 5330TCCCTCTCTCAGTTGCAATC 79 1763 1103743 N/A N/A 5412 5431TCTTTCTGTTTGTCTTTCAT 71 1764 1103759 N/A N/A 5482 5501GTGTCCTCTTCTGCCTGCCC 59 1765 1103775 N/A N/A 5775 5794TCTCTTGTACAGAGCAAGAA 110  1766 1103791 N/A N/A 5853 5872AGCCATCTCACTTCTCTGGT 67 1767 1103807 N/A N/A 5930 5949ACTACTAATAATAGCAATAG 82 1768 1103823 N/A N/A 5975 5994GTAATCCATGAAAAGCACAG 97 1769 1103839 N/A N/A 6022 6041CTTCGGCTCTCTCATCTGTC 76 1770 1103855 N/A N/A 6203 6222CAGGTCAGACACCTCTCTGT 76 1771 1103871 N/A N/A 6265 6284GCTCTGAGCTGTGGTGTTCT 88 1772 1103887 N/A N/A 6314 6333CCAAATGCCCCCTCTACAGT 92 1773 1103903 N/A N/A 6409 6428CCTCTCTGTGCTTTTCTGCC 60 1774 1103919 N/A N/A 6516 6535GGTCCTCCCAGCCCCATCGG 79 1775 1103935 N/A N/A 6906 6925CTTAGGCGAGCGGAGGCCTG 85 1776 1103951 N/A N/A 7021 7040GCCAGACCTCAGCACCTAGC 65 1777 1103967 N/A N/A 7316 7335GCTCACCGTGCCGCGCAGAG 76 1778 1103983 N/A N/A 7420 7439TCAGGCCCCGCCCTCGACCC 88 1779 1103999 N/A N/A 7806 7825AGATGAGCTCTACCGTGAGG 66 1780 1104015 N/A N/A 7930 7949ATCCATTCAGTCATCAAACA 90 1781 1104031 N/A N/A 8037 8056GTGAGGAGTCCAATCTTGGC 77 1782 1104047 N/A N/A 8076 8095GGTGAAGAAAGTTCCAAGGA 59 1783 1104063 N/A N/A 8229 8248TTTTCCTTGCCAGGAAAGTC 76 1784 1104079 N/A N/A 9360 9379CCCCCGCCCCGCCCGAGAGA 103  1785 1104095 N/A N/A 9413 9432CTAAGAATCATTTCAGGGCC 65 1786 1104111 N/A N/A 9454 9473TAAATATTCTAGTCCAGAAG 87 1787 1104127 N/A N/A 9508 9527GGTTAGCCTTTCTGATGCTG 30 1788 1104143 N/A N/A 9531 9550GTATTACCTCTACTAGTCAG 56 1789 1104159 N/A N/A 9558 9577TGACCTGTGACTATCTAGGA 29 1790 1104175 N/A N/A 9578 9597GCCATTTATCTGTGCTTTAG 10 1791 1104191 N/A N/A 9664 9683AAAGGCTGTTAAACATGTGG 39 1792 1104207 N/A N/A 9707 9726GCCTACTTCTCTAGGTGGGA 69 1793 1104223 N/A N/A 10557 10576ACGGTATTGAAATCAGGAGA 55 1794 1104239 N/A N/A 10639 10658TGGCTGCTCTGTCTTCTGGC 52 1795 1104255 N/A N/A 10748 10767CCCTCCCCATCATGAGTATG 79 1796 1104271 N/A N/A 10817 10836GAGGAGGCCTCTCATGGACT 95 1797 1104287 N/A N/A 11082 11101ACGGAATTACATTCAGTTTC 77 1798 1104303 N/A N/A 11151 11170ATTAGGATCCCATCTAGTGG 68 1799 1104319 N/A N/A 11229 11248AAGCTACTAACTTTAATTCT 92 1800 1104335 N/A N/A 11281 11300GAAATGGAAAGCCCTCCCCA 113  1801 1104351 N/A N/A 11389 11408GTGGTAAGCTGCTGGAGTAA 60 1802 1104367 N/A N/A 11498 11517CGACTTCCCAGGTCTTACTT 74 1803

TABLE 25 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  7 517 1103153 44 63 3492 3511GCTCTGGCTCTGCTCGCTCC 83 1804 1103169 354 373 3802 3821GGTTCAGCTCAGCAGCCAGC 85 1805 1103185 566 585 4951 4970TCTATAGGCAGCCAGGTTGT  73* 1806 1103201 762 781 5648 5667GGATCTCTTTCAGGGCTGCG 90 1807 1103217 1185 1204 N/A N/AGAATGGTGATCCGGTTCTCC 23 1808 1103233 1350 1369 11639 11658TGCCTCACATCACATCCTTG 98 1809 1103249 1528 1547 11817 11836CAGGCCTGATACTGACGGAG 103  1810 1103265 1670 1689 11959 11978AGTAGGTGCCCCCCGCCCTC 74 1811 1103281 1750 1769 12039 12058TTCCCAGATACTCCGAGAGA 65 1812 1103297 1861 1880 12150 12169ACCAAAAGACAAAACAAGCC 113  1813 1103313 1887 1906 12176 12195GCATAGGGATATCCCACCTC 95 1814 1103329 2097 2116 12386 12405TGAGTCATCGCTCAGGAGGT 47 1815 1103345 2331 2350 12620 12639TCCTCCTCCATCTCTACCAG 44 1816 1103361 2431 2450 12720 12739CAAGAGGATGAGTCACTTCC 49 1817 1103377 2485 2504 12774 12793AGTCATGCCCTGCCCCCATG 46 1818 1103393 2516 2535 12805 12824AGCAAGGAAGAGGCCTTTAG 68 1819 1103409 2673 2692 12962 12981TGTGTGTGAGTAAGAAGGGA 30 1820 1103425 2725 2744 13014 13033CTCAGTTTTACAATTGTAAA 71 1821 1103441 2905 2924 13194 13213GGGAGAGGAGAACCCTGAAG 98 1822 1103473 N/A N/A 8813 8832CTGGTGAGCCTGTATTGGTA 68 1823 1103489 N/A N/A 8975 8994AAAATGACGCAGTCCAGGCC 108  1824 1103505 N/A N/A 9046 9065TAACAAGCTCTGCCAGTTTA 92 1825 1103521 N/A N/A 9238 9257GAACTGAGTCAGCACTGAGC 124  1826 1103537 N/A N/A 9279 9298TATTCACTGCAAGAGCTGCG 111  1827 1103553 N/A N/A 9313 9332ATATTTAACATTAAGAGCAG 84 1828 1103569 N/A N/A 8533 8552CCTGGTATGATAGGCTCTGG 56 1829 1103585 N/A N/A 8655 8674ATCTGCAGACAGGGCAGATG 113  1830 1103601 N/A N/A 4001 4020AGCCCCTTCTGCTCACAAGG 74 1831 1103617 N/A N/A 4172 4191CCTGGCAGAAGGCATGCGGG 120  1832 1103633 N/A N/A 4295 4314CAGGCTCAGAATAGGTGAGC 80 1833 1103649 N/A N/A 4410 4429CCTGTGCTTTGCGCCCAGAC 92 1834 1103665 N/A N/A 4505 4524CCACCTTTTGAAATGAATTT 63 1835 1103681 N/A N/A 4641 4660ATCCACAAGCATACACTCAC 62 1836 1103697 N/A N/A 4845 4864TGAATACCTGCCTCAGTCTC 68 1837 1103713 N/A N/A 5101 5120CCCATTCCTCAGCCTTGCCT 95 1838 1103729 N/A N/A 5351 5370TGAGTGTCTCTCTCAGTCTC 132  1839 1103745 N/A N/A 5420 5439CCTTAGTGTCTTTCTGTTTG 69 1840 1103761 N/A N/A 5489 5508CGCCATTGTGTCCTCTTCTG 86 1841 1103777 N/A N/A 5778 5797CATTCTCTTGTACAGAGCAA 105  1842 1103793 N/A N/A 5861 5880CCTGGGCAAGCCATCTCACT 119  1843 1103809 N/A N/A 5943 5962AACAGTATCAGCTACTACTA 80 1844 1103825 N/A N/A 5979 5998TAAGGTAATCCATGAAAAGC 101  1845 1103841 N/A N/A 6108 6127CCCAAGTGAGATGTGCTAGA 118  1846 1103857 N/A N/A 6205 6224TCCAGGTCAGACACCTCTCT 97 1847 1103873 N/A N/A 6268 6287TAAGCTCTGAGCTGTGGTGT 51 1848 1103889 N/A N/A 6316 6335GCCCAAATGCCCCCTCTACA 73 1849 1103905 N/A N/A 6415 6434GCCCTGCCTCTCTGTGCTTT 64 1850 1103921 N/A N/A 6522 6541CAGGGAGGTCCTCCCAGCCC 101  1851 1103937 N/A N/A 6948 6967TTCCACACTGACAGCTGCAT 76 1852 1103953 N/A N/A 7077 7096TGGCTGGGACTTTTCCCAAC 113  1853 1103969 N/A N/A 7330 7349CCTGCCCTGGCCGCGCTCAC 87 1854 1103985 N/A N/A 7423 7442CGTTCAGGCCCCGCCCTCGA 78 1855 1104001 N/A N/A 7815 7834AAGGGAGCAAGATGAGCTCT 77 1856 1104017 N/A N/A 7956 7975CTAATCAATATTGGTTGAAT 87 1857 1104033 N/A N/A 8039 8058AGGTGAGGAGTCCAATCTTG 82 1858 1104049 N/A N/A 8082 8101GAAAGTGGTGAAGAAAGTTC 109  1859 1104065 N/A N/A 8252 8271CCATCATGACAACTTGAACG 110  1860 1104081 N/A N/A 9377 9396AATTTAGCTCCCCCCTCCCC 83 1861 1104097 N/A N/A 9417 9436TTAGCTAAGAATCATTTCAG 125  1862 1104113 N/A N/A 9456 9475CCTAAATATTCTAGTCCAGA 47 1863 1104129 N/A N/A 9511 9530CCTGGTTAGCCTTTCTGATG 47 1864 1104145 N/A N/A 9537 9556TTGGCAGTATTACCTCTACT 19 1865 1104161 N/A N/A 9561 9580TAGTGACCTGTGACTATCTA 47 1866 1104177 N/A N/A 9580 9599CTGCCATTTATCTGTGCTTT 52 1867 1104193 N/A N/A 9672 9691ACTAAAATAAAGGCTGTTAA 67 1868 1104209 N/A N/A 9786 9805AACCAGCCACATGACTCTGG 91 1869 1104225 N/A N/A 10559 10578GCACGGTATTGAAATCAGGA 30 1870 1104241 N/A N/A 10655 10674GCTGCTGCCAGAGTCCTGGC 69 1871 1104257 N/A N/A 10761 10780CCCCATCGCACCCCCCTCCC 103  1872 1104273 N/A N/A 10843 10862CAGGCTGGTTTCTGCAGATG 92 1873 1104289 N/A N/A 11084 11103AAACGGAATTACATTCAGTT 67 1874 1104305 N/A N/A 11153 11172GTATTAGGATCCCATCTAGT 58 1875 1104321 N/A N/A 11231 11250GAAAGCTACTAACTTTAATT 60 1876 1104337 N/A N/A 11308 11327CCCTGGTAGTTTCCTTTTAC 55 1877 1104353 N/A N/A 11392 11411CGAGTGGTAAGCTGCTGGAG 69 1878 1104369 N/A N/A 11512 11531CCTTGGGAAGTCCCCGACTT 78 1879

TABLE 26 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  6 517 1103154 109 128 3557 3576ATCATCTCCCCTGAGGAGAC 139  1880 1103170 376 395 3824 3843GTGGGCTCCTTGGCCCGCAG 89 1881 1103186 576 595 N/A N/ACTGCTTCCTGTCTATAGGCA  30* 1882 1103202 763 782 5649 5668CGGATCTCTTTCAGGGCTGC 38 1883 1103218 1202 1221 8351 8370GGAGAAGGTCTGCACGGGAA 43 1884 1103234 1355 1374 11644 11663GGTCCTGCCTCACATCACAT 87 1885 1103250 1532 1551 11821 11840CTGGCAGGCCTGATACTGAC 93 1886 1103266 1679 1698 11968 11987GGGCGATGTAGTAGGTGCCC 100  1887 1103282 1762 1781 12051 12070TCAAAGGCACAGTTCCCAGA 89 1888 1103298 1862 1881 12151 12170AACCAAAAGACAAAACAAGC 80 1889 1103314 1889 1908 12178 12197CAGCATAGGGATATCCCACC 86 1890 1103330 2098 2117 12387 12406TTGAGTCATCGCTCAGGAGG 49 1891 1103346 2332 2351 12621 12640CTCCTCCTCCATCTCTACCA 81 1892 1103362 2436 2455 12725 12744ATCTTCAAGAGGATGAGTCA 116  1893 1103378 2487 2506 12776 12795AAAGTCATGCCCTGCCCCCA 74 1894 1103394 2526 2545 12815 12834GGTATGACACAGCAAGGAAG 59 1895 1103410 2674 2693 12963 12982TTGTGTGTGAGTAAGAAGGG 52 1896 1103426 2730 2749 13019 13038CGTGCCTCAGTTTTACAATT 73 1897 1103442 2963 2982 13252 13271TGGAGGGAAAGGACACCAAG 79 1898 1103458 N/A N/A 8740 8759TCTTTGGTGCTTTTGCCCCC 50 1899 1103474 N/A N/A 8814 8833TCTGGTGAGCCTGTATTGGT 66 1900 1103490 N/A N/A 8978 8997GGGAAAATGACGCAGTCCAG 80 1901 1103506 N/A N/A 9076 9095GCGCACCCAAGGACTCACCA 115  1902 1103522 N/A N/A 9240 9259CTGAACTGAGTCAGCACTGA 90 1903 1103538 N/A N/A 9285 9304AAACTTTATTCACTGCAAGA 57 1904 1103570 N/A N/A 8537 8556GTACCCTGGTATGATAGGCT 22 1905 1103586 N/A N/A 8684 8703ACACTCAGAAGGGCAGTGCT 115  1906 1103602 N/A N/A 4002 4021CAGCCCCTTCTGCTCACAAG 77 1907 1103618 N/A N/A 4181 4200CCCTGGACTCCTGGCAGAAG 98 1908 1103634 N/A N/A 4336 4355CATCAACCTTCTCCGCTTCC 75 1909 1103650 N/A N/A 4411 4430ACCTGTGCTTTGCGCCCAGA 92 1910 1103666 N/A N/A 4566 4585TGCATGTCCTGTCAGCTCAG 122  1911 1103682 N/A N/A 4645 4664GCGCATCCACAAGCATACAC 113  1912 1103698 N/A N/A 4846 4865TTGAATACCTGCCTCAGTCT 104  1913 1103714 N/A N/A 5140 5159TGAGGAGTAGAGGGCCACTG 64 1914 1103730 N/A N/A 5353 5372TCTGAGTGTCTCTCTCAGTC 135  1915 1103746 N/A N/A 5422 5441TCCCTTAGTGTCTTTCTGTT 101  1916 1103762 N/A N/A 5494 5513CCCCACGCCATTGTGTCCTC 85 1917 1103778 N/A N/A 5780 5799CCCATTCTCTTGTACAGAGC 72 1918 1103794 N/A N/A 5862 5881CCCTGGGCAAGCCATCTCAC 80 1919 1103810 N/A N/A 5948 5967AACAGAACAGTATCAGCTAC 82 1920 1103826 N/A N/A 5980 5999GTAAGGTAATCCATGAAAAG 73 1921 1103842 N/A N/A 6112 6131GGCTCCCAAGTGAGATGTGC 118  1922 1103858 N/A N/A 6207 6226CTTCCAGGTCAGACACCTCT 75 1923 1103874 N/A N/A 6269 6288ATAAGCTCTGAGCTGTGGTG 50 1924 1103890 N/A N/A 6317 6336AGCCCAAATGCCCCCTCTAC 77 1925 1103906 N/A N/A 6449 6468CAGCTAGGTGCCCTGGCTAG 131  1926 1103922 N/A N/A 6531 6550CCCTGCCTGCAGGGAGGTCC 69 1927 1103938 N/A N/A 6970 6989CAGTGCCACAGAATCCAGAA 94 1928 1103954 N/A N/A 7104 7123CCCCAGTTAACCCCAGGACG 130  1929 1103970 N/A N/A 7337 7356TCCCCGTCCTGCCCTGGCCG 65 1930 1103986 N/A N/A 7454 7473GCCCCAGGCCCCGCCTCTAG 174  1931 1104002 N/A N/A 7824 7843CAGTCCCTGAAGGGAGCAAG 113  1932 1104018 N/A N/A 7958 7977GCCTAATCAATATTGGTTGA 71 1933 1104034 N/A N/A 8041 8060TGAGGTGAGGAGTCCAATCT 69 1934 1104050 N/A N/A 8107 8126CTGAAGGAAGATGGAAAAGG 78 1935 1104066 N/A N/A 8254 8273GGCCATCATGACAACTTGAA 105  1936 1104082 N/A N/A 9378 9397TAATTTAGCTCCCCCCTCCC 106  1937 1104098 N/A N/A 9418 9437CTTAGCTAAGAATCATTTCA 96 1938 1104114 N/A N/A 9457 9476TCCTAAATATTCTAGTCCAG 63 1939 1104130 N/A N/A 9513 9532AGCCTGGTTAGCCTTTCTGA 68 1940 1104146 N/A N/A 9538 9557TTTGGCAGTATTACCTCTAC 73 1941 1104162 N/A N/A 9562 9581TTAGTGACCTGTGACTATCT 92 1942 1104178 N/A N/A 9582 9601CTCTGCCATTTATCTGTGCT 48 1943 1104194 N/A N/A 9675 9694TTAACTAAAATAAAGGCTGT 132  1944 1104210 N/A N/A 9810 9829TCAAGAAGTCCCAACTTAGC 74 1945 1104226 N/A N/A 10560 10579AGCACGGTATTGAAATCAGG 77 1946 1104242 N/A N/A 10656 10675TGCTGCTGCCAGAGTCCTGG 89 1947 1104258 N/A N/A 10773 10792CATGCCCGGCTTCCCCATCG 98 1948 1104274 N/A N/A 10923 10942CTTACCTCTCCATCCCGCAT 100  1949 1104290 N/A N/A 11088 11107GAGGAAACGGAATTACATTC 95 1950 1104306 N/A N/A 11154 11173TGTATTAGGATCCCATCTAG 135  1951 1104322 N/A N/A 11232 11251TGAAAGCTACTAACTTTAAT 75 1952 1104338 N/A N/A 11312 11331ATTCCCCTGGTAGTTTCCTT 97 1953 1104354 N/A N/A 11393 11412GCGAGTGGTAAGCTGCTGGA 57 1954 1104370 N/A N/A 11523 11542AGGCTGTGTAACCTTGGGAA 75 1955

TABLE 27 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  7 517 1103155 110 129 3558 3577CATCATCTCCCCTGAGGAGA 90 1956 1103171 380 399 3828 3847CTTGGTGGGCTCCTTGGCCC 103  1957 1103187 577 596 N/A N/ATCTGCTTCCTGTCTATAGGC  33* 1958 1103203 764 783 5650 5669GCGGATCTCTTTCAGGGCTG 65 1959 1103219 1220 1239 N/A N/ATTCTCGAATCTGCAGGTTGG 111  1960 1103235 1361 1380 11650 11669CAGGTGGGTCCTGCCTCACA 147  1961 1103251 1533 1552 11822 11841TCTGGCAGGCCTGATACTGA 134  1962 1103267 1681 1700 11970 11989GAGGGCGATGTAGTAGGTGC 66 1963 1103283 1768 1787 12057 12076GGAAACTCAAAGGCACAGTT 72 1964 1103299 1870 1889 12159 12178CTCATAAAAACCAAAAGACA 128  1965 1103315 1890 1909 12179 12198GCAGCATAGGGATATCCCAC 89 1966 1103331 2107 2126 12396 12415CTGAGACACTTGAGTCATCG 45 1967 1103347 2334 2353 12623 12642GCCTCCTCCTCCATCTCTAC 53 1968 1103363 2440 2459 12729 12748CAGCATCTTCAAGAGGATGA 109  1969 1103379 2488 2507 12777 12796CAAAGTCATGCCCTGCCCCC 94 1970 1103395 2527 2546 12816 12835TGGTATGACACAGCAAGGAA 48 1971 1103411 2676 2695 12965 12984TTTTGTGTGTGAGTAAGAAG 43 1972 1103427 2750 2769 13039 13058GCCAGTGTCTTCACTTTGCT 73 1973 1103443 3013 3032 13302 13321CAGTGCCCTGAAGATTAGCA 68 1974 1103459 N/A N/A 8741 8760GTCTTTGGTGCTTTTGCCCC 51 1975 1103475 N/A N/A 8815 8834ATCTGGTGAGCCTGTATTGG 45 1976 1103491 N/A N/A 8979 8998TGGGAAAATGACGCAGTCCA 61 1977 1103507 N/A N/A 9091 9110CAGAGCAGCTCCACTGCGCA 127  1978 1103523 N/A N/A 9241 9260TCTGAACTGAGTCAGCACTG 84 1979 1103539 N/A N/A 9286 9305AAAACTTTATTCACTGCAAG 64 1980 1103555 N/A N/A 8369 8388ACCTCGAATCTGCAGGTTGG 130  1981 1103571 N/A N/A 8538 8557AGTACCCTGGTATGATAGGC 33 1982 1103587 N/A N/A 8714 8733GACAAGCAGTTAAAAAAACA 62 1983 1103603 N/A N/A 4003 4022TCAGCCCCTTCTGCTCACAA 58 1984 1103619 N/A N/A 4210 4229CTTTCCCCAGTAGGGAGGTG 86 1985 1103635 N/A N/A 4343 4362TCATGGACATCAACCTTCTC 87 1986 1103651 N/A N/A 4414 4433GTCACCTGTGCTTTGCGCCC 107  1987 1103667 N/A N/A 4568 4587CATGCATGTCCTGTCAGCTC 72 1988 1103683 N/A N/A 4646 4665GGCGCATCCACAAGCATACA 112  1989 1103699 N/A N/A 4855 4874TGAGGACACTTGAATACCTG 67 1990 1103715 N/A N/A 5163 5182GCTTCCTGGAGTGGCAGGAG 109* 1991 1103731 N/A N/A 5362 5381TCTCCTCTCTCTGAGTGTCT 69 1992 1103747 N/A N/A 5424 5443TCTCCCTTAGTGTCTTTCTG 65 1993 1103763 N/A N/A 5499 5518GGGTTCCCCACGCCATTGTG 109  1994 1103779 N/A N/A 5782 5801CCCCCATTCTCTTGTACAGA 136  1995 1103795 N/A N/A 5881 5900AGCAATAGTGCCTGTGTGAC 69 1996 1103811 N/A N/A 5949 5968GAACAGAACAGTATCAGCTA 91 1997 1103827 N/A N/A 5981 6000AGTAAGGTAATCCATGAAAA 109  1998 1103843 N/A N/A 6113 6132AGGCTCCCAAGTGAGATGTG 89 1999 1103859 N/A N/A 6214 6233ATCACACCTTCCAGGTCAGA 68 2000 1103875 N/A N/A 6271 6290ATATAAGCTCTGAGCTGTGG 87 2001 1103891 N/A N/A 6318 6337TAGCCCAAATGCCCCCTCTA 101  2002 1103907 N/A N/A 6454 6473CTCCTCAGCTAGGTGCCCTG 66 2003 1103923 N/A N/A 6534 6553CTCCCCTGCCTGCAGGGAGG 124  2004 1103939 N/A N/A 6975 6994ATGAACAGTGCCACAGAATC 124  2005 1103955 N/A N/A 7105 7124ACCCCAGTTAACCCCAGGAC 90 2006 1103971 N/A N/A 7339 7358CGTCCCCGTCCTGCCCTGGC 93 2007 1103987 N/A N/A 7473 7492TTTCGAGGCCCGGCCCCCGG 78 2008 1104003 N/A N/A 7829 7848AGACTCAGTCCCTGAAGGGA 89 2009 1104019 N/A N/A 7959 7978GGCCTAATCAATATTGGTTG 80 2010 1104035 N/A N/A 8043 8062GGTGAGGTGAGGAGTCCAAT 62 2011 1104051 N/A N/A 8116 8135CATGTCTATCTGAAGGAAGA 159  2012 1104067 N/A N/A 8283 8302GGACACCCCTAGGCTGGGTC 121  2013 1104083 N/A N/A 9384 9403AAAAAGTAATTTAGCTCCCC 108  2014 1104099 N/A N/A 9419 9438CCTTAGCTAAGAATCATTTC 76 2015 1104115 N/A N/A 9458 9477GTCCTAAATATTCTAGTCCA 55 2016 1104131 N/A N/A 9514 9533CAGCCTGGTTAGCCTTTCTG 38 2017 1104147 N/A N/A 9539 9558ATTTGGCAGTATTACCTCTA 51 2018 1104163 N/A N/A 9563 9582TTTAGTGACCTGTGACTATC 108  2019 1104179 N/A N/A 9583 9602TCTCTGCCATTTATCTGTGC 48 2020 1104195 N/A N/A 9676 9695ATTAACTAAAATAAAGGCTG 90 2021 1104211 N/A N/A 9811 9830CTCAAGAAGTCCCAACTTAG 83 2022 1104227 N/A N/A 10562 10581CCAGCACGGTATTGAAATCA 50 2023 1104243 N/A N/A 10659 10678TAGTGCTGCTGCCAGAGTCC 54 2024 1104259 N/A N/A 10777 10796CTCCCATGCCCGGCTTCCCC 147  2025 1104275 N/A N/A 10981 11000TGGCCTGGCCTTGAGAATCC 108  2026 1104291 N/A N/A 11100 11119TATGGAGGGACTGAGGAAAC 107  2027 1104307 N/A N/A 11155 11174GTGTATTAGGATCCCATCTA 19 2028 1104323 N/A N/A 11233 11252GTGAAAGCTACTAACTTTAA 51 2029 1104339 N/A N/A 11325 11344ATCCCCTCTTCCCATTCCCC 128  2030 1104355 N/A N/A 11394 11413GGCGAGTGGTAAGCTGCTGG 99 2031 1104371 N/A N/A 11525 11544CGAGGCTGTGTAACCTTGGG 99 2032

TABLE 28 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG 14 517 1103156 142 161 3590 3609CCCAGACGGCGGCCAGGAGC 113  2033 1103172 408 427 3856 3875GCAGCTCAGCCTGGTAGACG 92 2034 1103188 582 601 N/A N/ACTTCATCTGCTTCCTGTCTA  23* 2035 1103204 829 848 N/A N/AAACTTGGAGCGGTACCACTC 106  2036 1103220 1249 1268 10863 10882TCTGACACAGACTTGGTGTC 110  2037 1103236 1397 1416 11686 11705CTGCTCGGGCCCCTCATGAG 18 2038 1103252 1534 1553 11823 11842GTCTGGCAGGCCTGATACTG 84 2039 1103268 1682 1701 11971 11990GGAGGGCGATGTAGTAGGTG 31 2040 1103284 1798 1817 12087 12106CTGAGTCTCAGTTTTCCTCC 48 2041 1103300 1871 1890 12160 12179CCTCATAAAAACCAAAAGAC 76 2042 1103316 1891 1910 12180 12199GGCAGCATAGGGATATCCCA 74 2043 1103332 2108 2127 12397 12416ACTGAGACACTTGAGTCATC 77 2044 1103348 2339 2358 12628 12647CAATTGCCTCCTCCTCCATC 91 2045 1103364 2444 2463 12733 12752GTTTCAGCATCTTCAAGAGG 63 2046 1103380 2489 2508 12778 12797ACAAAGTCATGCCCTGCCCC 89 2047 1103396 2528 2547 12817 12836CTGGTATGACACAGCAAGGA 67 2048 1103412 2677 2696 12966 12985ATTTTGTGTGTGAGTAAGAA 49 2049 1103428 2752 2771 13041 13060GAGCCAGTGTCTTCACTTTG 38 2050 1103444 3014 3033 13303 13322GCAGTGCCCTGAAGATTAGC 88 2051 1103460 N/A N/A 8746 8765TCCCCGTCTTTGGTGCTTTT 74 2052 1103476 N/A N/A 8823 8842CATTTACAATCTGGTGAGCC 103  2053 1103492 N/A N/A 8981 9000CCTGGGAAAATGACGCAGTC 88 2054 1103508 N/A N/A 9103 9122GCTCAGAGGCCCCAGAGCAG 117  2055 1103524 N/A N/A 9242 9261CTCTGAACTGAGTCAGCACT 93 2056 1103540 N/A N/A 9288 9307ATAAAACTTTATTCACTGCA 82 2057 1103556 N/A N/A 8397 8416GCCCTTCCCACGAGGCCCTG 102  2058 1103572 N/A N/A 8540 8559GAAGTACCCTGGTATGATAG 80 2059 1103588 N/A N/A 8715 8734TGACAAGCAGTTAAAAAAAC 98 2060 1103604 N/A N/A 4004 4023TTCAGCCCCTTCTGCTCACA 97 2061 1103620 N/A N/A 4212 4231TGCTTTCCCCAGTAGGGAGG 52 2062 1103636 N/A N/A 4351 4370AATCTCCCTCATGGACATCA 102  2063 1103652 N/A N/A 4421 4440GCAGGCAGTCACCTGTGCTT 84 2064 1103668 N/A N/A 4576 4595AAGGCACACATGCATGTCCT 103  2065 1103684 N/A N/A 4663 4682CTGCTTCTGCTCACACAGGC 109  2066 1103700 N/A N/A 4856 4875CTGAGGACACTTGAATACCT 120  2067 1103716 N/A N/A 5165 5184CTGCTTCCTGGAGTGGCAGG 123* 2068 1103732 N/A N/A 5363 5382CTCTCCTCTCTCTGAGTGTC 100  2069 1103748 N/A N/A 5432 5451TCTTTCCGTCTCCCTTAGTG 94 2070 1103764 N/A N/A 5518 5537AGGGTACAGGCCACAGCTGG 101  2071 1103780 N/A N/A 5785 5804CTTCCCCCATTCTCTTGTAC 106  2072 1103796 N/A N/A 5883 5902AAAGCAATAGTGCCTGTGTG 80 2073 1103812 N/A N/A 5950 5969AGAACAGAACAGTATCAGCT 96 2074 1103828 N/A N/A 5982 6001TAGTAAGGTAATCCATGAAA 110  2075 1103844 N/A N/A 6126 6145AGGCATGGAATGCAGGCTCC 103  2076 1103860 N/A N/A 6215 6234TATCACACCTTCCAGGTCAG 102  2077 1103876 N/A N/A 6272 6291CATATAAGCTCTGAGCTGTG 91 2078 1103892 N/A N/A 6319 6338CTAGCCCAAATGCCCCCTCT 103  2079 1103908 N/A N/A 6456 6475CACTCCTCAGCTAGGTGCCC 104  2080 1103924 N/A N/A 6535 6554TCTCCCCTGCCTGCAGGGAG 111  2081 1103940 N/A N/A 6977 6996GAATGAACAGTGCCACAGAA 94 2082 1103956 N/A N/A 7106 7125CACCCCAGTTAACCCCAGGA 102  2083 1103972 N/A N/A 7340 7359CCGTCCCCGTCCTGCCCTGG 90 2084 1103988 N/A N/A 7474 7493GTTTCGAGGCCCGGCCCCCG 117  2085 1104004 N/A N/A 7831 7850AAAGACTCAGTCCCTGAAGG 119  2086 1104020 N/A N/A 7960 7979AGGCCTAATCAATATTGGTT 112  2087 1104036 N/A N/A 8044 8063GGGTGAGGTGAGGAGTCCAA 68 2088 1104052 N/A N/A 8117 8136GCATGTCTATCTGAAGGAAG 94 2089 1104068 N/A N/A 8289 8308TGCCTGGGACACCCCTAGGC 122  2090 1104084 N/A N/A 9385 9404TAAAAAGTAATTTAGCTCCC 91 2091 1104100 N/A N/A 9427 9446ATTTTGTTCCTTAGCTAAGA 96 2092 1104116 N/A N/A 9459 9478GGTCCTAAATATTCTAGTCC 35 2093 1104132 N/A N/A 9515 9534TCAGCCTGGTTAGCCTTTCT 37 2094 1104148 N/A N/A 9540 9559GATTTGGCAGTATTACCTCT 52 2095 1104164 N/A N/A 9564 9583CTTTAGTGACCTGTGACTAT 84 2096 1104180 N/A N/A 9588 9607ACTTCTCTCTGCCATTTATC 110  2097 1104196 N/A N/A 9677 9696AATTAACTAAAATAAAGGCT 108  2098 1104212 N/A N/A 9820 9839AGACCAGAGCTCAAGAAGTC 119  2099 1104228 N/A N/A 10563 10582CCCAGCACGGTATTGAAATC 84 2100 1104244 N/A N/A 10660 10679ATAGTGCTGCTGCCAGAGTC 88 2101 1104260 N/A N/A 10779 10798CTCTCCCATGCCCGGCTTCC 112  2102 1104276 N/A N/A 10983 11002CATGGCCTGGCCTTGAGAAT 31 2103 1104292 N/A N/A 11115 11134CCCCTTAGAACAGCCTATGG 111  2104 1104308 N/A N/A 11156 11175TGTGTATTAGGATCCCATCT 41 2105 1104324 N/A N/A 11234 11253AGTGAAAGCTACTAACTTTA 78 2106 1104340 N/A N/A 11326 11345AATCCCCTCTTCCCATTCCC 105  2107 1104356 N/A N/A 11396 11415CTGGCGAGTGGTAAGCTGCT 91 2108 1104372 N/A N/A 11535 11554TCCACCACCACGAGGCTGTG 101  2109

TABLE 29 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  8 517 1103157 158 177 3606 3625GAGGCGGGTGCCAGGACCCA 45 2110 1103173 409 428 3857 3876CGCAGCTCAGCCTGGTAGAC 97 2111 1103189 636 655 5225 5244TCTCCTCCTCCAGCGACTCA 92 2112 1103205 843 862 N/A N/ACTGTCAGGTCTGCAAACTTG 74 2113 1103221 1263 1282 10877 10896TCTTGAGGTGGCCTTCTGAC 104  2114 1103237 1399 1418 11688 11707TTCTGCTCGGGCCCCTCATG 75 2115 1103253 1544 1563 11833 11852GGTGGGTGCCGTCTGGCAGG 34 2116 1103269 1683 1702 11972 11991TGGAGGGCGATGTAGTAGGT 58 2117 1103285 1800 1819 12089 12108GTCTGAGTCTCAGTTTTCCT 25 2118 1103301 1872 1891 12161 12180ACCTCATAAAAACCAAAAGA 116  2119 1103317 1893 1912 12182 12201TAGGCAGCATAGGGATATCC 101  2120 1103333 2110 2129 12399 12418GGACTGAGACACTTGAGTCA 96 2121 1103349 2348 2367 12637 12656GCGCCATCCCAATTGCCTCC 98 2122 1103365 2446 2465 12735 12754CTGTTTCAGCATCTTCAAGA 41 2123 1103381 2493 2512 12782 12801TGGGACAAAGTCATGCCCTG 105  2124 1103397 2530 2549 12819 12838GCCTGGTATGACACAGCAAG 85 2125 1103413 2697 2716 12986 13005CTACCTAGAATACTGGGTAC 110  2126 1103429 2753 2772 13042 13061TGAGCCAGTGTCTTCACTTT 70 2127 1103445 3015 3034 13304 13323AGCAGTGCCCTGAAGATTAG 86 2128 1103461 N/A N/A 8748 8767TTTCCCCGTCTTTGGTGCTT 77 2129 1103477 N/A N/A 8824 8843CCATTTACAATCTGGTGAGC 61 2130 1103493 N/A N/A 8982 9001TCCTGGGAAAATGACGCAGT 114  2131 1103509 N/A N/A 9142 9161TCCCAGTGACAGGAAGAGGT 105  2132 1103525 N/A N/A 9245 9264ATCCTCTGAACTGAGTCAGC 93 2133 1103541 N/A N/A 9292 9311GAACATAAAACTTTATTCAC 110  2134 1103557 N/A N/A 8403 8422TCCAGTGCCCTTCCCACGAG 104  2135 1103573 N/A N/A 8542 8561TAGAAGTACCCTGGTATGAT 110  2136 1103589 N/A N/A 8728 8747TTGCCCCCTGTAGTGACAAG 103  2137 1103605 N/A N/A 4005 4024ATTCAGCCCCTTCTGCTCAC 106  2138 1103621 N/A N/A 4213 4232CTGCTTTCCCCAGTAGGGAG 94 2139 1103637 N/A N/A 4367 4386CTTCACCCCAGAATCCAATC 124  2140 1103653 N/A N/A 4423 4442TGGCAGGCAGTCACCTGTGC 130  2141 1103669 N/A N/A 4577 4596GAAGGCACACATGCATGTCC 103  2142 1103685 N/A N/A 4665 4684ACCTGCTTCTGCTCACACAG 96 2143 1103701 N/A N/A 4858 4877TTCTGAGGACACTTGAATAC 128  2144 1103717 N/A N/A 5166 5185TCTGCTTCCTGGAGTGGCAG 111* 2145 1103733 N/A N/A 5364 5383TCTCTCCTCTCTCTGAGTGT 100  2146 1103749 N/A N/A 5440 5459CTCTTGTCTCTTTCCGTCTC 105  2147 1103765 N/A N/A 5535 5554AAGCGGTACCAGGGCTCAGG 62 2148 1103781 N/A N/A 5810 5829TATTCTCCCAGCTTCCTCCA 112  2149 1103797 N/A N/A 5888 5907TTTTGAAAGCAATAGTGCCT 94 2150 1103813 N/A N/A 5951 5970TAGAACAGAACAGTATCAGC 92 2151 1103829 N/A N/A 5989 6008TACTTCATAGTAAGGTAATC 112  2152 1103845 N/A N/A 6144 6163CCCAAAACAGACTGGCAGAG 79 2153 1103861 N/A N/A 6216 6235ATATCACACCTTCCAGGTCA 87 2154 1103877 N/A N/A 6273 6292ACATATAAGCTCTGAGCTGT 105  2155 1103893 N/A N/A 6333 6352CGTGCCTGTCCTGCCTAGCC 90 2156 1103909 N/A N/A 6457 6476ACACTCCTCAGCTAGGTGCC 87 2157 1103925 N/A N/A 6536 6555TTCTCCCCTGCCTGCAGGGA 120  2158 1103941 N/A N/A 6978 6997TGAATGAACAGTGCCACAGA 112  2159 1103957 N/A N/A 7107 7126GCACCCCAGTTAACCCCAGG 93 2160 1103973 N/A N/A 7341 7360CCCGTCCCCGTCCTGCCCTG 98 2161 1103989 N/A N/A 7476 7495AGGTTTCGAGGCCCGGCCCC 93 2162 1104005 N/A N/A 7851 7870GATCTGCACACAAGGCTGAA 99 2163 1104021 N/A N/A 7961 7980GAGGCCTAATCAATATTGGT 111  2164 1104037 N/A N/A 8047 8066GATGGGTGAGGTGAGGAGTC 40 2165 1104053 N/A N/A 8118 8137CGCATGTCTATCTGAAGGAA 90 2166 1104069 N/A N/A 8325 8344TCCTGAAAGAAAGCAGAGGG 114  2167 1104085 N/A N/A 9386 9405GTAAAAAGTAATTTAGCTCC 127  2168 1104101 N/A N/A 9428 9447AATTTTGTTCCTTAGCTAAG 125  2169 1104117 N/A N/A 9460 9479TGGTCCTAAATATTCTAGTC 74 2170 1104133 N/A N/A 9516 9535GTCAGCCTGGTTAGCCTTTC 30 2171 1104149 N/A N/A 9541 9560GGATTTGGCAGTATTACCTC 52 2172 1104165 N/A N/A 9565 9584GCTTTAGTGACCTGTGACTA 43 2173 1104181 N/A N/A 9589 9608TACTTCTCTCTGCCATTTAT 75 2174 1104197 N/A N/A 9678 9697CAATTAACTAAAATAAAGGC 130  2175 1104213 N/A N/A 9828 9847GGATCAGGAGACCAGAGCTC 118  2176 1104229 N/A N/A 10564 10583ACCCAGCACGGTATTGAAAT 105  2177 1104245 N/A N/A 10670 10689CCAAATCCCAATAGTGCTGC 80 2178 1104261 N/A N/A 10782 10801ATCCTCTCCCATGCCCGGCT 87 2179 1104277 N/A N/A 11006 11025TGTGCCAGCCCCAGGCTTTC 125  2180 1104293 N/A N/A 11119 11138GGCTCCCCTTAGAACAGCCT 115  2181 1104309 N/A N/A 11158 11177AGTGTGTATTAGGATCCCAT 18 2182 1104325 N/A N/A 11235 11254AAGTGAAAGCTACTAACTTT 102  2183 1104341 N/A N/A 11327 11346AAATCCCCTCTTCCCATTCC 110  2184 1104357 N/A N/A 11428 11447AGATAACACTGGGAAAGCAT 97 2185 1104373 N/A N/A 11537 11556GGTCCACCACCACGAGGCTG 108  2186

TABLE 30 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  7 517 1103158 206 225 3654 3673GGAGAAATCCACCCGGGTCG 155  2187 1103174 410 429 3858 3877TCGCAGCTCAGCCTGGTAGA 127  2188 1103190 648 667 5237 5256TCAAGAACCGGATCTCCTCC 63 2189 1103206 926 945 7283 7302GGTCAAGGACTGCAACTGGC 96 2190 1103222 1270 1289 10884 10903ATGTTCCTCTTGAGGTGGCC 86 2191 1103238 1400 1419 11689 11708CTTCTGCTCGGGCCCCTCAT 59 2192 1103254 1573 1592 11862 11881TTCTTGTTAGTTGGAGTTGC 33 2193 1103270 1686 1705 11975 11994ATGTGGAGGGCGATGTAGTA 49 2194 1103286 1809 1828 12098 12117CCCTTTCCTGTCTGAGTCTC 72 2195 1103302 1873 1892 12162 12181CACCTCATAAAAACCAAAAG 100  2196 1103318 1996 2015 12285 12304GAACAACCCTCTGAGCTGGG 57 2197 1103334 2124 2143 12413 12432ATGGCAGCTCAGGTGGACTG 54 2198 1103350 2364 2383 12653 12672CCTACTTGTATGCCTAGCGC 75 2199 1103366 2447 2466 12736 12755CCTGTTTCAGCATCTTCAAG 42 2200 1103382 2494 2513 12783 12802ATGGGACAAAGTCATGCCCT 98 2201 1103398 2567 2586 12856 12875AAGAAGCAGCAGTCCCAGGG 75 2202 1103414 2700 2719 12989 13008GCACTACCTAGAATACTGGG 76 2203 1103430 2754 2773 13043 13062ATGAGCCAGTGTCTTCACTT 37 2204 1103446 3017 3036 13306 13325GCAGCAGTGCCCTGAAGATT 29 2205 1103462 N/A N/A 8749 8768TTTTCCCCGTCTTTGGTGCT 56 2206 1103478 N/A N/A 8825 8844TCCATTTACAATCTGGTGAG 41 2207 1103494 N/A N/A 8983 9002TTCCTGGGAAAATGACGCAG 97 2208 1103510 N/A N/A 9144 9163CCTCCCAGTGACAGGAAGAG 104  2209 1103526 N/A N/A 9246 9265AATCCTCTGAACTGAGTCAG 54 2210 1103542 N/A N/A 9293 9312GGAACATAAAACTTTATTCA 69 2211 1103558 N/A N/A 8405 8424ACTCCAGTGCCCTTCCCACG 27 2212 1103574 N/A N/A 8543 8562CTAGAAGTACCCTGGTATGA 95 2213 1103590 N/A N/A 8730 8749TTTTGCCCCCTGTAGTGACA 67 2214 1103606 N/A N/A 4038 4057TCCCTGAGACTTCTCGGGCA 110  2215 1103622 N/A N/A 4214 4233ACTGCTTTCCCCAGTAGGGA 77 2216 1103638 N/A N/A 4371 4390CTTTCTTCACCCCAGAATCC 139  2217 1103654 N/A N/A 4424 4443GTGGCAGGCAGTCACCTGTG 83 2218 1103670 N/A N/A 4578 4597TGAAGGCACACATGCATGTC 105  2219 1103686 N/A N/A 4666 4685CACCTGCTTCTGCTCACACA 158  2220 1103702 N/A N/A 4960 4979CCTGACCTGTCTATAGGCAG  92* 2221 1103718 N/A N/A 5167 5186ATCTGCTTCCTGGAGTGGCA 102* 2222 1103734 N/A N/A 5365 5384TTCTCTCCTCTCTCTGAGTG 102  2223 1103750 N/A N/A 5442 5461CTCTCTTGTCTCTTTCCGTC 69 2224 1103766 N/A N/A 5536 5555GAAGCGGTACCAGGGCTCAG 95 2225 1103782 N/A N/A 5811 5830ATATTCTCCCAGCTTCCTCC 92 2226 1103798 N/A N/A 5893 5912GGTACTTTTGAAAGCAATAG 47 2227 1103814 N/A N/A 5955 5974GGCTTAGAACAGAACAGTAT 87 2228 1103830 N/A N/A 5996 6015CAGCACCTACTTCATAGTAA 54 2229 1103846 N/A N/A 6156 6175GAGGCTCAGTAACCCAAAAC 60 2230 1103862 N/A N/A 6217 6236AATATCACACCTTCCAGGTC 130  2231 1103878 N/A N/A 6275 6294CTACATATAAGCTCTGAGCT 127  2232 1103894 N/A N/A 6369 6388AGGAATGGCCCTCCCTTCTT 93 2233 1103910 N/A N/A 6458 6477CACACTCCTCAGCTAGGTGC 75 2234 1103926 N/A N/A 6541 6560GCCCTTTCTCCCCTGCCTGC 106  2235 1103942 N/A N/A 6980 6999AATGAATGAACAGTGCCACA 113  2236 1103958 N/A N/A 7114 7133GTGAGCAGCACCCCAGTTAA 117  2237 1103974 N/A N/A 7345 7364TCCGCCCGTCCCCGTCCTGC 119  2238 1103990 N/A N/A 7477 7496GAGGTTTCGAGGCCCGGCCC 129  2239 1104006 N/A N/A 7852 7871GGATCTGCACACAAGGCTGA 112  2240 1104022 N/A N/A 7962 7981TGAGGCCTAATCAATATTGG 96 2241 1104038 N/A N/A 8050 8069AGAGATGGGTGAGGTGAGGA 36 2242 1104054 N/A N/A 8140 8159CTTGAGTGTTATCTGGGAGG 61 2243 1104070 N/A N/A 8336 8355GGGAATGGTGATCCTGAAAG 53 2244 1104086 N/A N/A 9388 9407AAGTAAAAAGTAATTTAGCT 87 2245 1104102 N/A N/A 9429 9448GAATTTTGTTCCTTAGCTAA 80 2246 1104118 N/A N/A 9477 9496GAGACATGCATATCTAGTGG 23 2247 1104134 N/A N/A 9518 9537TAGTCAGCCTGGTTAGCCTT 63 2248 1104150 N/A N/A 9547 9566TATCTAGGATTTGGCAGTAT 58 2249 1104166 N/A N/A 9567 9586GTGCTTTAGTGACCTGTGAC 25 2250 1104182 N/A N/A 9591 9610CCTACTTCTCTCTGCCATTT 75 2251 1104198 N/A N/A 9679 9698ACAATTAACTAAAATAAAGG 95 2252 1104214 N/A N/A 10003 10022GTAATCCCCTTACTCGGGAG 84 2253 1104230 N/A N/A 10565 10584AACCCAGCACGGTATTGAAA 117  2254 1104246 N/A N/A 10671 10690CCCAAATCCCAATAGTGCTG 59 2255 1104262 N/A N/A 10786 10805AGCCATCCTCTCCCATGCCC 130  2256 1104278 N/A N/A 11010 11029TCTATGTGCCAGCCCCAGGC 83 2257 1104294 N/A N/A 11120 11139AGGCTCCCCTTAGAACAGCC 75 2258 1104310 N/A N/A 11159 11178GAGTGTGTATTAGGATCCCA 18 2259 1104326 N/A N/A 11237 11256GGAAGTGAAAGCTACTAACT 46 2260 1104342 N/A N/A 11329 11348CCAAATCCCCTCTTCCCATT 81 2261 1104358 N/A N/A 11430 11449TGAGATAACACTGGGAAAGC 89 2262 1104374 N/A N/A 11543 11562ACCCCAGGTCCACCACCACG 101  2263

TABLE 31 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  7 517 1103159 218 237 3666 3685TGCCCCAGCCAGGGAGAAAT 93 2264 1103175 411 430 3859 3878CTCGCAGCTCAGCCTGGTAG 69 2265 1103191 652 671 5241 5260TTCCTCAAGAACCGGATCTC 79 2266 1103207 930 949 7287 7306CGCAGGTCAAGGACTGCAAC 84 2267 1103223 1271 1290 10885 10904GATGTTCCTCTTGAGGTGGC 94 2268 1103239 1401 1420 11690 11709GCTTCTGCTCGGGCCCCTCA 52 2269 1103255 1575 1594 11864 11883GTTTCTTGTTAGTTGGAGTT 19 2270 1103271 1687 1706 11976 11995GATGTGGAGGGCGATGTAGT 69 2271 1103287 1811 1830 12100 12119TTCCCTTTCCTGTCTGAGTC 56 2272 1103303 1874 1893 12163 12182CCACCTCATAAAAACCAAAA 89 2273 1103319 1997 2016 12286 12305AGAACAACCCTCTGAGCTGG 61 2274 1103335 2183 2202 12472 12491CAAGTGCTGAGAATCAAGCT 87 2275 1103351 2398 2417 12687 12706TCCAGAGGCCAAGTGCAACT 57 2276 1103367 2448 2467 12737 12756TCCTGTTTCAGCATCTTCAA 61 2277 1103383 2495 2514 12784 12803AATGGGACAAAGTCATGCCC 88 2278 1103399 2629 2648 12918 12937AAAAGCAGCCGGTCACTATG 100  2279 1103415 2703 2722 12992 13011AGGGCACTACCTAGAATACT 71 2280 1103431 2789 2808 13078 13097CAGCCCTGAGCACCCGGCCT 106  2281 1103447 3019 3038 13308 13327CAGCAGCAGTGCCCTGAAGA 38 2282 1103463 N/A N/A 8750 8769ATTTTCCCCGTCTTTGGTGC 65 2283 1103479 N/A N/A 8855 8874GGCAGGCAGCTAACCGCGAG 80 2284 1103495 N/A N/A 8984 9003GTTCCTGGGAAAATGACGCA 89 2285 1103511 N/A N/A 9146 9165GCCCTCCCAGTGACAGGAAG 96 2286 1103527 N/A N/A 9249 9268AAGAATCCTCTGAACTGAGT 85 2287 1103543 N/A N/A 9294 9313GGGAACATAAAACTTTATTC 42 2288 1103559 N/A N/A 8407 8426GGACTCCAGTGCCCTTCCCA 90 2289 1103575 N/A N/A 8544 8563CCTAGAAGTACCCTGGTATG 64 2290 1103591 N/A N/A 8732 8751GCTTTTGCCCCCTGTAGTGA 36 2291 1103607 N/A N/A 4039 4058CTCCCTGAGACTTCTCGGGC 129  2292 1103623 N/A N/A 4216 4235GCACTGCTTTCCCCAGTAGG 88 2293 1103639 N/A N/A 4372 4391ACTTTCTTCACCCCAGAATC 101  2294 1103655 N/A N/A 4456 4475AGTCAAAGTAACTTGATGGG 90 2295 1103671 N/A N/A 4579 4598TTGAAGGCACACATGCATGT 102  2296 1103687 N/A N/A 4728 4747AGGATGAGCAGATGTGGGCT 77 2297 1103703 N/A N/A 4961 4980CCCTGACCTGTCTATAGGCA 105* 2298 1103719 N/A N/A 5169 5188TCATCTGCTTCCTGGAGTGG  49* 2299 1103735 N/A N/A 5366 5385TTTCTCTCCTCTCTCTGAGT 95 2300 1103751 N/A N/A 5446 5465AATGCTCTCTTGTCTCTTTC 83 2301 1103767 N/A N/A 5537 5556AGAAGCGGTACCAGGGCTCA 100  2302 1103783 N/A N/A 5830 5849AGTCAGTCACCTGGAGAGGA 79 2303 1103799 N/A N/A 5898 5917TAATGGGTACTTTTGAAAGC 100  2304 1103815 N/A N/A 5956 5975GGGCTTAGAACAGAACAGTA 74 2305 1103831 N/A N/A 5997 6016ACAGCACCTACTTCATAGTA 113  2306 1103847 N/A N/A 6158 6177TAGAGGCTCAGTAACCCAAA 74 2307 1103863 N/A N/A 6218 6237CAATATCACACCTTCCAGGT 72 2308 1103879 N/A N/A 6277 6296CACTACATATAAGCTCTGAG 93 2309 1103895 N/A N/A 6370 6389TAGGAATGGCCCTCCCTTCT 91 2310 1103911 N/A N/A 6461 6480ACTCACACTCCTCAGCTAGG 100  2311 1103927 N/A N/A 6546 6565TAATAGCCCTTTCTCCCCTG 93 2312 1103943 N/A N/A 6982 7001CGAATGAATGAACAGTGCCA 108  2313 1103959 N/A N/A 7120 7139AGGGAGGTGAGCAGCACCCC 103  2314 1103975 N/A N/A 7347 7366GCTCCGCCCGTCCCCGTCCT 66 2315 1103991 N/A N/A 7478 7497GGAGGTTTCGAGGCCCGGCC 117  2316 1104007 N/A N/A 7853 7872GGGATCTGCACACAAGGCTG 49 2317 1104023 N/A N/A 7963 7982TTGAGGCCTAATCAATATTG 110  2318 1104039 N/A N/A 8052 8071GAAGAGATGGGTGAGGTGAG 36 2319 1104055 N/A N/A 8193 8212CTTTTTCCCCAGCAGCCAAC 125  2320 1104071 N/A N/A 9325 9344TCACATTCACTAATATTTAA 74 2321 1104087 N/A N/A 9389 9408CAAGTAAAAAGTAATTTAGC 120  2322 1104103 N/A N/A 9433 9452AGAGGAATTTTGTTCCTTAG 98 2323 1104119 N/A N/A 9481 9500TCCTGAGACATGCATATCTA 82 2324 1104135 N/A N/A 9520 9539ACTAGTCAGCCTGGTTAGCC 66 2325 1104151 N/A N/A 9549 9568ACTATCTAGGATTTGGCAGT 36 2326 1104167 N/A N/A 9569 9588CTGTGCTTTAGTGACCTGTG 42 2327 1104183 N/A N/A 9636 9655AGTGCTCAATACACATAGGT 58 2328 1104199 N/A N/A 9680 9699GACAATTAACTAAAATAAAG 87 2329 1104215 N/A N/A 10131 10150ACTCCGTCGAAAGCAGGCAA 91 2330 1104231 N/A N/A 10566 10585AAACCCAGCACGGTATTGAA 71 2331 1104247 N/A N/A 10694 10713TCTTCCAAACGGGCTGGAGA 98 2332 1104263 N/A N/A 10790 10809CATGAGCCATCCTCTCCCAT 70 2333 1104279 N/A N/A 11023 11042TTGCTGGGAACCTTCTATGT 97 2334 1104295 N/A N/A 11121 11140AAGGCTCCCCTTAGAACAGC 64 2335 1104311 N/A N/A 11160 11179AGAGTGTGTATTAGGATCCC 31 2336 1104327 N/A N/A 11242 11261GAGTTGGAAGTGAAAGCTAC 67 2337 1104343 N/A N/A 11342 11361CGTGGCGGATACGCCAAATC 96 2338 1104359 N/A N/A 11431 11450GTGAGATAACACTGGGAAAG 65 2339 1104375 N/A N/A 11552 11571TTCACACAGACCCCAGGTCC 96 2340

TABLE 32 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  6 517 1103160 221 240 3669 3688GAGTGCCCCAGCCAGGGAGA 119  2341 1103176 442 461 3890 3909GCGGTGAGTTGATCGAGCCG 86 2342 1103192 653 672 5242 5261CTTCCTCAAGAACCGGATCT 81 2343 1103208 932 951 7289 7308GTCGCAGGTCAAGGACTGCA 100  2344 1103224 1293 1312 10907 10926GCATCTCCACGGTCTTCACC 85 2345 1103240 1403 1422 11692 11711CTGCTTCTGCTCGGGCCCCT 80 2346 1103256 1577 1596 11866 11885GAGTTTCTTGTTAGTTGGAG  9 2347 1103272 1700 1719 11989 12008AACAGGAATCAGGGATGTGG 35 2348 1103288 1836 1855 12125 12144CCAGGGCTACCTTGTCTGTG 90 2349 1103304 1878 1897 12167 12186TATCCCACCTCATAAAAACC 88 2350 1103320 2001 2020 12290 12309TAGGAGAACAACCCTCTGAG 62 2351 1103336 2185 2204 12474 12493CCCAAGTGCTGAGAATCAAG 53 2352 1103352 2400 2419 12689 12708AATCCAGAGGCCAAGTGCAA 95 2353 1103368 2452 2471 12741 12760TCTCTCCTGTTTCAGCATCT 22 2354 1103384 2496 2515 12785 12804AAATGGGACAAAGTCATGCC 98 2355 1103400 2637 2656 12926 12945GCTTAGGGAAAAGCAGCCGG 51 2356 1103416 2704 2723 12993 13012TAGGGCACTACCTAGAATAC 75 2357 1103432 2791 2810 13080 13099GTCAGCCCTGAGCACCCGGC 116  2358 1103448 3021 3040 13310 13329GGCAGCAGCAGTGCCCTGAA 38 2359 1103464 N/A N/A 8752 8771TGATTTTCCCCGTCTTTGGT 58 2360 1103480 N/A N/A 8866 8885CGTGTCTGAGAGGCAGGCAG 76 2361 1103496 N/A N/A 8989 9008CTGCAGTTCCTGGGAAAATG 70 2362 1103512 N/A N/A 9147 9166GGCCCTCCCAGTGACAGGAA 68 2363 1103528 N/A N/A 9250 9269GAAGAATCCTCTGAACTGAG 64 2364 1103544 N/A N/A 9295 9314AGGGAACATAAAACTTTATT 57 2365 1103560 N/A N/A 8408 8427AGGACTCCAGTGCCCTTCCC 57 2366 1103576 N/A N/A 8546 8565CACCTAGAAGTACCCTGGTA 101  2367 1103592 N/A N/A 8734 8753GTGCTTTTGCCCCCTGTAGT 43 2368 1103608 N/A N/A 4040 4059CCTCCCTGAGACTTCTCGGG 86 2369 1103624 N/A N/A 4217 4236TGCACTGCTTTCCCCAGTAG 102  2370 1103640 N/A N/A 4373 4392CACTTTCTTCACCCCAGAAT 135  2371 1103656 N/A N/A 4465 4484AGCTGTGCAAGTCAAAGTAA 80 2372 1103672 N/A N/A 4584 4603TGCACTTGAAGGCACACATG 59 2373 1103688 N/A N/A 4729 4748GAGGATGAGCAGATGTGGGC 65 2374 1103704 N/A N/A 4962 4981TCCCTGACCTGTCTATAGGC  95* 2375 1103720 N/A N/A 5170 5189TTCATCTGCTTCCTGGAGTG  72* 2376 1103736 N/A N/A 5381 5400ACCTGCCAATCTCTGTTTCT 94 2377 1103752 N/A N/A 5447 5466GAATGCTCTCTTGTCTCTTT 54 2378 1103768 N/A N/A 5541 5560TGAGAGAAGCGGTACCAGGG 79 2379 1103784 N/A N/A 5832 5851AAAGTCAGTCACCTGGAGAG 87 2380 1103800 N/A N/A 5907 5926CAGTAATAATAATGGGTACT 67 2381 1103816 N/A N/A 5957 5976AGGGCTTAGAACAGAACAGT 50 2382 1103832 N/A N/A 6005 6024GTCAAAGAACAGCACCTACT 121  2383 1103848 N/A N/A 6160 6179GGTAGAGGCTCAGTAACCCA 68 2384 1103864 N/A N/A 6219 6238TCAATATCACACCTTCCAGG 80 2385 1103880 N/A N/A 6278 6297ACACTACATATAAGCTCTGA 109  2386 1103896 N/A N/A 6382 6401TCTGTCCTCCACTAGGAATG 120  2387 1103912 N/A N/A 6462 6481CACTCACACTCCTCAGCTAG 85 2388 1103928 N/A N/A 6554 6573CTGGGTTCTAATAGCCCTTT 77 2389 1103944 N/A N/A 6983 7002GCGAATGAATGAACAGTGCC 75 2390 1103960 N/A N/A 7122 7141TCAGGGAGGTGAGCAGCACC 111  2391 1103976 N/A N/A 7354 7373CGTCCCTGCTCCGCCCGTCC 93 2392 1103992 N/A N/A 7504 7523TCCCGCGGAGCCCCGACCCG 90 2393 1104008 N/A N/A 7857 7876GGAAGGGATCTGCACACAAG 59 2394 1104024 N/A N/A 7965 7984CCTTGAGGCCTAATCAATAT 80 2395 1104040 N/A N/A 8053 8072AGAAGAGATGGGTGAGGTGA 85 2396 1104056 N/A N/A 8201 8220TCTCCTAGCTTTTTCCCCAG 131  2397 1104072 N/A N/A 9327 9346CGTCACATTCACTAATATTT 22 2398 1104088 N/A N/A 9395 9414CCAATGCAAGTAAAAAGTAA 92 2399 1104104 N/A N/A 9434 9453AAGAGGAATTTTGTTCCTTA 72 2400 1104120 N/A N/A 9483 9502AGTCCTGAGACATGCATATC 77 2401 1104136 N/A N/A 9521 9540TACTAGTCAGCCTGGTTAGC 69 2402 1104152 N/A N/A 9550 9569GACTATCTAGGATTTGGCAG 22 2403 1104168 N/A N/A 9570 9589TCTGTGCTTTAGTGACCTGT 56 2404 1104184 N/A N/A 9637 9656TAGTGCTCAATACACATAGG 58 2405 1104200 N/A N/A 9681 9700AGACAATTAACTAAAATAAA 85 2406 1104216 N/A N/A 10132 10151GACTCCGTCGAAAGCAGGCA 63 2407 1104232 N/A N/A 10570 10589CTGAAAACCCAGCACGGTAT 142  2408 1104248 N/A N/A 10695 10714CTCTTCCAAACGGGCTGGAG 69 2409 1104264 N/A N/A 10791 10810GCATGAGCCATCCTCTCCCA 121  2410 1104280 N/A N/A 11024 11043GTTGCTGGGAACCTTCTATG 34 2411 1104296 N/A N/A 11122 11141CAAGGCTCCCCTTAGAACAG 56 2412 1104312 N/A N/A 11162 11181AGAGAGTGTGTATTAGGATC 37 2413 1104328 N/A N/A 11243 11262AGAGTTGGAAGTGAAAGCTA 73 2414 1104344 N/A N/A 11370 11389AGATGAGCTCCCACTGTGGT 74 2415 1104360 N/A N/A 11432 11451GGTGAGATAACACTGGGAAA 48 2416 1104376 N/A N/A 11553 11572GTTCACACAGACCCCAGGTC 96 2417

TABLE 33 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  5 517 1103161 222 241 3670 3689TGAGTGCCCCAGCCAGGGAG 168  2418 1103177 443 462 3891 3910GGCGGTGAGTTGATCGAGCC 101  2419 1103193 654 673 5243 5262TCTTCCTCAAGAACCGGATC 68 2420 1103209 948 967 7305 7324CGCGCAGAGACTCCAGGTCG 112  2421 1103225 1295 1314 10909 10928CCGCATCTCCACGGTCTTCA 74 2422 1103241 1409 1428 11698 11717ACTATCCTGCTTCTGCTCGG 41 2423 1103257 1578 1597 11867 11886TGAGTTTCTTGTTAGTTGGA 24 2424 1103273 1702 1721 11991 12010ACAACAGGAATCAGGGATGT 94 2425 1103289 1840 1859 12129 12148CTGGCCAGGGCTACCTTGTC 95 2426 1103305 1879 1898 12168 12187ATATCCCACCTCATAAAAAC 108  2427 1103321 2013 2032 12302 12321TCAGGGTCAGTCTAGGAGAA 63 2428 1103337 2189 2208 12478 12497ATCCCCCAAGTGCTGAGAAT 96 2429 1103353 2403 2422 12692 12711CACAATCCAGAGGCCAAGTG 70 2430 1103369 2453 2472 12742 12761TTCTCTCCTGTTTCAGCATC 32 2431 1103385 2497 2516 12786 12805GAAATGGGACAAAGTCATGC 79 2432 1103401 2638 2657 12927 12946GGCTTAGGGAAAAGCAGCCG 140  2433 1103417 2705 2724 12994 13013ATAGGGCACTACCTAGAATA 117  2434 1103433 2792 2811 13081 13100TGTCAGCCCTGAGCACCCGG 64 2435 1103449 3031 3050 13320 13339AGCGACTAAAGGCAGCAGCA 64 2436 1103465 N/A N/A 8753 8772GTGATTTTCCCCGTCTTTGG 26 2437 1103481 N/A N/A 8892 8911ACTCCCTGTCAAGCTGGGCA 110  2438 1103497 N/A N/A 8991 9010CACTGCAGTTCCTGGGAAAA 92 2439 1103513 N/A N/A 9170 9189TCCAGGCACAGCGAGACCCA 121  2440 1103529 N/A N/A 9251 9270GGAAGAATCCTCTGAACTGA 86 2441 1103545 N/A N/A 9296 9315CAGGGAACATAAAACTTTAT 135  2442 1103561 N/A N/A 8409 8428CAGGACTCCAGTGCCCTTCC 97 2443 1103577 N/A N/A 8548 8567CCCACCTAGAAGTACCCTGG 72 2444 1103593 N/A N/A 8736 8755TGGTGCTTTTGCCCCCTGTA 84 2445 1103609 N/A N/A 4044 4063GGCCCCTCCCTGAGACTTCT 161  2446 1103625 N/A N/A 4218 4237CTGCACTGCTTTCCCCAGTA 123  2447 1103641 N/A N/A 4381 4400CTTTCCCTCACTTTCTTCAC 135  2448 1103657 N/A N/A 4479 4498GACCACCGCTTCACAGCTGT 118  2449 1103673 N/A N/A 4594 4613CATGTCCTCCTGCACTTGAA 66 2450 1103689 N/A N/A 4745 4764ACAGAGGACTTGTCTGGAGG 77 2451 1103705 N/A N/A 4963 4982CTCCCTGACCTGTCTATAGG  97* 2452 1103721 N/A N/A 5171 5190CTTCATCTGCTTCCTGGAGT  78* 2453 1103737 N/A N/A 5382 5401TACCTGCCAATCTCTGTTTC 79 2454 1103753 N/A N/A 5449 5468TCGAATGCTCTCTTGTCTCT 92 2455 1103769 N/A N/A 5543 5562GGTGAGAGAAGCGGTACCAG 141  2456 1103785 N/A N/A 5834 5853TGAAAGTCAGTCACCTGGAG 102  2457 1103801 N/A N/A 5908 5927GCAGTAATAATAATGGGTAC 69 2458 1103817 N/A N/A 5958 5977CAGGGCTTAGAACAGAACAG 61 2459 1103833 N/A N/A 6014 6033CTCTCATCTGTCAAAGAACA 87 2460 1103849 N/A N/A 6161 6180TGGTAGAGGCTCAGTAACCC 60 2461 1103865 N/A N/A 6246 6265TCTACGGGCACTATGTTTGG 79 2462 1103881 N/A N/A 6279 6298CACACTACATATAAGCTCTG 188  2463 1103897 N/A N/A 6398 6417TTTTCTGCCTCCAGGCTCTG 93 2464 1103913 N/A N/A 6469 6488CTTCTGCCACTCACACTCCT 101  2465 1103929 N/A N/A 6806 6825GTCAGTGGCACAATCCCGGG 76 2466 1103945 N/A N/A 6987 7006GCAAGCGAATGAATGAACAG 76 2467 1103961 N/A N/A 7123 7142ATCAGGGAGGTGAGCAGCAC 51 2468 1103977 N/A N/A 7360 7379GGTGGCCGTCCCTGCTCCGC 89 2469 1103993 N/A N/A 7760 7779TGATGAGGGCTCACCGGTTC 77 2470 1104009 N/A N/A 7896 7915GCTATGTGTGAGGCAGGCAC 138  2471 1104025 N/A N/A 7979 7998ACCCAAGTCCTTGGCCTTGA 79 2472 1104041 N/A N/A 8054 8073CAGAAGAGATGGGTGAGGTG 71 2473 1104057 N/A N/A 8202 8221ATCTCCTAGCTTTTTCCCCA 147  2474 1104073 N/A N/A 9345 9364AGAGAGAAAAATATAACACG 90 2475 1104089 N/A N/A 9401 9420TCAGGGCCAATGCAAGTAAA 53 2476 1104105 N/A N/A 9436 9455AGAAGAGGAATTTTGTTCCT 68 2477 1104121 N/A N/A 9484 9503AAGTCCTGAGACATGCATAT 64 2478 1104137 N/A N/A 9522 9541CTACTAGTCAGCCTGGTTAG 68 2479 1104153 N/A N/A 9551 9570TGACTATCTAGGATTTGGCA 38 2480 1104169 N/A N/A 9571 9590ATCTGTGCTTTAGTGACCTG 80 2481 1104185 N/A N/A 9638 9657TTAGTGCTCAATACACATAG 75 2482 1104201 N/A N/A 9682 9701GAGACAATTAACTAAAATAA 86 2483 1104217 N/A N/A 10133 10152AGACTCCGTCGAAAGCAGGC 78 2484 1104233 N/A N/A 10571 10590TCTGAAAACCCAGCACGGTA 116  2485 1104249 N/A N/A 10696 10715GCTCTTCCAAACGGGCTGGA 116  2486 1104265 N/A N/A 10795 10814CAGGGCATGAGCCATCCTCT 101  2487 1104281 N/A N/A 11056 11075CTCCTCCAGAATTCCCTGGG 140  2488 1104297 N/A N/A 11130 11149TTTGGTACCAAGGCTCCCCT 72 2489 1104313 N/A N/A 11178 11197TTTGAGGGTGAGAAAGAGAG 82 2490 1104329 N/A N/A 11259 11278CCCAACTGTGTCTGCTAGAG 47 2491 1104345 N/A N/A 11371 11390AAGATGAGCTCCCACTGTGG 111  2492 1104361 N/A N/A 11434 11453GTGGTGAGATAACACTGGGA 43 2493 1104377 N/A N/A 11557 11576AGGAGTTCACACAGACCCCA 92 2494

TABLE 34 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  6 517 1103162 224 243 3672 3691ATTGAGTGCCCCAGCCAGGG 94 2495 1103178 468 487 3916 3935TCTCAACCTCCAGCCGGGCG  23* 2496 1103194 687 706 5573 5592GCTCCTGGAGTTCCCGAACC 139  2497 1103210 949 968 7306 7325CCGCGCAGAGACTCCAGGTC 98 2498 1103226 1316 1335 N/A N/ACTCCTTAATGACCTCTCCAT 95 2499 1103242 1419 1438 11708 11727AGGCGGAGCAACTATCCTGC 150  2500 1103258 1579 1598 11868 11887GTGAGTTTCTTGTTAGTTGG 35 2501 1103274 1706 1725 11995 12014CATAACAACAGGAATCAGGG 49 2502 1103290 1846 1865 12135 12154AAGCCTCTGGCCAGGGCTAC 113  2503 1103306 1880 1899 12169 12188GATATCCCACCTCATAAAAA 98 2504 1103322 2015 2034 12304 12323GATCAGGGTCAGTCTAGGAG 46 2505 1103338 2196 2215 12485 12504ACAACAGATCCCCCAAGTGC 89 2506 1103354 2405 2424 12694 12713CCCACAATCCAGAGGCCAAG 79 2507 1103370 2454 2473 12743 12762TTTCTCTCCTGTTTCAGCAT 31 2508 1103386 2498 2517 12787 12806AGAAATGGGACAAAGTCATG 102  2509 1103402 2643 2662 12932 12951CCCTTGGCTTAGGGAAAAGC 102  2510 1103418 2706 2725 12995 13014AATAGGGCACTACCTAGAAT 130  2511 1103434 2793 2812 13082 13101GTGTCAGCCCTGAGCACCCG 73 2512 1103450 3032 3051 13321 13340CAGCGACTAAAGGCAGCAGC 76 2513 1103466 N/A N/A 8759 8778GACCTTGTGATTTTCCCCGT 55 2514 1103482 N/A N/A 8893 8912CACTCCCTGTCAAGCTGGGC 112  2515 1103498 N/A N/A 9028 9047TAATGTACAGTTACTCTGTA 80 2516 1103514 N/A N/A 9206 9225CCTCAGGGATGAAAGAATAA 56 2517 1103530 N/A N/A 9252 9271GGGAAGAATCCTCTGAACTG 83 2518 1103546 N/A N/A 9297 9316GCAGGGAACATAAAACTTTA 132  2519 1103562 N/A N/A 8441 8460CTGGAGCAACCTACAGGCCC 103  2520 1103578 N/A N/A 8550 8569GCCCCACCTAGAAGTACCCT 83 2521 1103594 N/A N/A 8738 8757TTTGGTGCTTTTGCCCCCTG 76 2522 1103610 N/A N/A 4045 4064CGGCCCCTCCCTGAGACTTC 92 2523 1103626 N/A N/A 4219 4238CCTGCACTGCTTTCCCCAGT 124  2524 1103642 N/A N/A 4384 4403GCTCTTTCCCTCACTTTCTT 111  2525 1103658 N/A N/A 4480 4499TGACCACCGCTTCACAGCTG 66 2526 1103674 N/A N/A 4598 4617CGCACATGTCCTCCTGCACT 122  2527 1103690 N/A N/A 4746 4765GACAGAGGACTTGTCTGGAG 69 2528 1103706 N/A N/A 4966 4985CACCTCCCTGACCTGTCTAT 165* 2529 1103722 N/A N/A 5173 5192GGCTTCATCTGCTTCCTGGA  11* 2530 1103738 N/A N/A 5390 5409CCTGTCTCTACCTGCCAATC 93 2531 1103754 N/A N/A 5456 5475CAGGAGTTCGAATGCTCTCT 77 2532 1103770 N/A N/A 5555 5574CCTCCTGACCAGGGTGAGAG 91 2533 1103786 N/A N/A 5838 5857CTGGTGAAAGTCAGTCACCT 84 2534 1103802 N/A N/A 5909 5928AGCAGTAATAATAATGGGTA 122  2535 1103818 N/A N/A 5960 5979CACAGGGCTTAGAACAGAAC 101  2536 1103834 N/A N/A 6015 6034TCTCTCATCTGTCAAAGAAC 81 2537 1103850 N/A N/A 6162 6181ATGGTAGAGGCTCAGTAACC 66 2538 1103866 N/A N/A 6247 6266CTCTACGGGCACTATGTTTG 72 2539 1103882 N/A N/A 6286 6305TGGCTCCCACACTACATATA 89 2540 1103898 N/A N/A 6401 6420TGCTTTTCTGCCTCCAGGCT 82 2541 1103914 N/A N/A 6476 6495CCAGTGGCTTCTGCCACTCA 92 2542 1103930 N/A N/A 6898 6917AGCGGAGGCCTGGGTGTTTT 74 2543 1103946 N/A N/A 6988 7007AGCAAGCGAATGAATGAACA 116  2544 1103962 N/A N/A 7124 7143AATCAGGGAGGTGAGCAGCA 71 2545 1103978 N/A N/A 7384 7403CCCTTCTCCCCTGGCATCTC 130  2546 1103994 N/A N/A 7792 7811GTGAGGCAGCAGGGAGACTT 80 2547 1104010 N/A N/A 7904 7923GACTGCCTGCTATGTGTGAG 118  2548 1104026 N/A N/A 7981 8000TGACCCAAGTCCTTGGCCTT 109  2549 1104042 N/A N/A 8056 8075GGCAGAAGAGATGGGTGAGG 75 2550 1104058 N/A N/A 8207 8226ACTCCATCTCCTAGCTTTTT 95 2551 1104074 N/A N/A 9347 9366CGAGAGAGAAAAATATAACA 129  2552 1104090 N/A N/A 9403 9422TTTCAGGGCCAATGCAAGTA 49 2553 1104106 N/A N/A 9447 9466TCTAGTCCAGAAGAAGAGGA 76 2554 1104122 N/A N/A 9494 9513ATGCTGAATTAAGTCCTGAG 38 2555 1104138 N/A N/A 9523 9542TCTACTAGTCAGCCTGGTTA 68 2556 1104154 N/A N/A 9553 9572TGTGACTATCTAGGATTTGG 68 2557 1104170 N/A N/A 9572 9591TATCTGTGCTTTAGTGACCT 53 2558 1104186 N/A N/A 9639 9658ATTAGTGCTCAATACACATA 101  2559 1104202 N/A N/A 9683 9702GGAGACAATTAACTAAAATA 94 2560 1104218 N/A N/A 10135 10154TAAGACTCCGTCGAAAGCAG 131  2561 1104234 N/A N/A 10572 10591TTCTGAAAACCCAGCACGGT 110  2562 1104250 N/A N/A 10708 10727GAGAGAGCCTAGGCTCTTCC 70 2563 1104266 N/A N/A 10796 10815TCAGGGCATGAGCCATCCTC 60 2564 1104282 N/A N/A 11065 11084TTCCTTGCTCTCCTCCAGAA 156  2565 1104298 N/A N/A 11144 11163TCCCATCTAGTGGCTTTGGT 48 2566 1104314 N/A N/A 11212 11231TCTCTTTCTCTCCCTGGCAA 74 2567 1104330 N/A N/A 11264 11283CCATCCCCAACTGTGTCTGC 64 2568 1104346 N/A N/A 11381 11400CTGCTGGAGTAAGATGAGCT 99 2569 1104362 N/A N/A 11478 11497TTCTTGATAGTAACCACAGC 53 2570 1104378 N/A N/A 11558 11577TAGGAGTTCACACAGACCCC 164  2571

TABLE 35 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  8 517 1103163 233 252 3681 3700GAAGCCAGCATTGAGTGCCC 109  2572 1103179 484 503 3932 3951TGTGCCAGATTGTCCCTCTC  13* 2573 1103195 713 732 5599 5618CACATGGACCTGCTGTCGGG 107  2574 1103211 951 970 7308 7327TGCCGCGCAGAGACTCCAGG 90 2575 1103227 1317 1336 N/A N/AACTCCTTAATGACCTCTCCA 151  2576 1103243 1434 1453 11723 11742GAAATGTGCCAGCAGAGGCG 96 2577 1103259 1581 1600 11870 11889GGGTGAGTTTCTTGTTAGTT 15 2578 1103275 1712 1731 12001 12020AGTTTCCATAACAACAGGAA 91 2579 1103291 1851 1870 12140 12159AAAACAAGCCTCTGGCCAGG 115  2580 1103307 1881 1900 12170 12189GGATATCCCACCTCATAAAA 107  2581 1103323 2030 2049 12319 12338CACCCATCTTAGACTGATCA 116  2582 1103339 2197 2216 12486 12505CACAACAGATCCCCCAAGTG 110  2583 1103355 2409 2428 12698 12717AATTCCCACAATCCAGAGGC 72 2584 1103371 2455 2474 12744 12763CTTTCTCTCCTGTTTCAGCA 20 2585 1103387 2506 2525 12795 12814AGGCCTTTAGAAATGGGACA 76 2586 1103403 2656 2675 12945 12964GGACCGCAAGAGGCCCTTGG 83 2587 1103419 2707 2726 12996 13015AAATAGGGCACTACCTAGAA 100  2588 1103435 2831 2850 13120 13139CTGCTCAGTCAAAGCAGAGT 99 2589 1103467 N/A N/A 8765 8784TCTTGTGACCTTGTGATTTT 55 2590 1103483 N/A N/A 8894 8913TCACTCCCTGTCAAGCTGGG 130  2591 1103499 N/A N/A 9029 9048TTAATGTACAGTTACTCTGT 98 2592 1103515 N/A N/A 9214 9233CGATGGAGCCTCAGGGATGA 57 2593 1103531 N/A N/A 9253 9272AGGGAAGAATCCTCTGAACT 59 2594 1103547 N/A N/A 9298 9317AGCAGGGAACATAAAACTTT 79 2595 1103563 N/A N/A 8458 8477TGATCCTCAGTCCCAGTCTG 95 2596 1103579 N/A N/A 8552 8571AAGCCCCACCTAGAAGTACC 131  2597 1103595 N/A N/A 3965 3984CCTCCTCACTTCTGCCTCAC  77* 2598 1103611 N/A N/A 4046 4065TCGGCCCCTCCCTGAGACTT 74 2599 1103627 N/A N/A 4220 4239TCCTGCACTGCTTTCCCCAG 80 2600 1103643 N/A N/A 4385 4404TGCTCTTTCCCTCACTTTCT 119  2601 1103659 N/A N/A 4484 4503ATTATGACCACCGCTTCACA 54 2602 1103675 N/A N/A 4603 4622ACACACGCACATGTCCTCCT 157  2603 1103691 N/A N/A 4751 4770CCTTAGACAGAGGACTTGTC 90 2604 1103707 N/A N/A 4967 4986CCACCTCCCTGACCTGTCTA 102* 2605 1103723 N/A N/A 5301 5320AGTTGCAATCTCTGTGTTGA 105  2606 1103739 N/A N/A 5391 5410TCCTGTCTCTACCTGCCAAT 81 2607 1103755 N/A N/A 5457 5476CCAGGAGTTCGAATGCTCTC 97 2608 1103771 N/A N/A 5557 5576AACCTCCTGACCAGGGTGAG 87 2609 1103787 N/A N/A 5839 5858TCTGGTGAAAGTCAGTCACC 99 2610 1103803 N/A N/A 5910 5929TAGCAGTAATAATAATGGGT 53 2611 1103819 N/A N/A 5967 5986TGAAAAGCACAGGGCTTAGA 118  2612 1103835 N/A N/A 6016 6035CTCTCTCATCTGTCAAAGAA 64 2613 1103851 N/A N/A 6164 6183ATATGGTAGAGGCTCAGTAA 70 2614 1103867 N/A N/A 6253 6272GGTGTTCTCTACGGGCACTA 87 2615 1103883 N/A N/A 6289 6308TCCTGGCTCCCACACTACAT 109  2616 1103899 N/A N/A 6402 6421GTGCTTTTCTGCCTCCAGGC 32 2617 1103915 N/A N/A 6489 6508CCCTGCTCAGACACCAGTGG 157  2618 1103931 N/A N/A 6899 6918GAGCGGAGGCCTGGGTGTTT 123  2619 1103947 N/A N/A 7005 7024TAGCACAACACCTGGTCAGC 92 2620 1103963 N/A N/A 7127 7146GGAAATCAGGGAGGTGAGCA 58 2621 1103979 N/A N/A 7385 7404GCCCTTCTCCCCTGGCATCT 83 2622 1103995 N/A N/A 7799 7818CTCTACCGTGAGGCAGCAGG 97 2623 1104011 N/A N/A 7909 7928CTAGTGACTGCCTGCTATGT 112  2624 1104027 N/A N/A 8007 8026GGGAGTATGCCTCTTAGTTT 55 2625 1104043 N/A N/A 8071 8090AGAAAGTTCCAAGGAGGCAG 112  2626 1104059 N/A N/A 8208 8227AACTCCATCTCCTAGCTTTT 85 2627 1104075 N/A N/A 9348 9367CCGAGAGAGAAAAATATAAC 66 2628 1104091 N/A N/A 9404 9423ATTTCAGGGCCAATGCAAGT 85 2629 1104107 N/A N/A 9448 9467TTCTAGTCCAGAAGAAGAGG 72 2630 1104123 N/A N/A 9495 9514GATGCTGAATTAAGTCCTGA 41 2631 1104139 N/A N/A 9527 9546TACCTCTACTAGTCAGCCTG 78 2632 1104155 N/A N/A 9554 9573CTGTGACTATCTAGGATTTG 70 2633 1104171 N/A N/A 9573 9592TTATCTGTGCTTTAGTGACC 54 2634 1104187 N/A N/A 9642 9661CATATTAGTGCTCAATACAC 59 2635 1104203 N/A N/A 9703 9722ACTTCTCTAGGTGGGAGAGA 58 2636 1104219 N/A N/A 10136 10155GTAAGACTCCGTCGAAAGCA 99 2637 1104235 N/A N/A 10573 10592TTTCTGAAAACCCAGCACGG 132  2638 1104251 N/A N/A 10730 10749TGAGAACCTATGCAACCGAG 62 2639 1104267 N/A N/A 10798 10817TTTCAGGGCATGAGCCATCC 140  2640 1104283 N/A N/A 11070 11089TCAGTTTCCTTGCTCTCCTC 82 2641 1104299 N/A N/A 11146 11165GATCCCATCTAGTGGCTTTG 39 2642 1104315 N/A N/A 11213 11232TTCTCTTTCTCTCCCTGGCA 85 2643 1104331 N/A N/A 11265 11284CCCATCCCCAACTGTGTCTG 73 2644 1104347 N/A N/A 11384 11403AAGCTGCTGGAGTAAGATGA 122  2645 1104363 N/A N/A 11483 11502TACTTTTCTTGATAGTAACC 74 2646 1104379 N/A N/A 11559 11578TTAGGAGTTCACACAGACCC 85 2647

TABLE 36 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG 10 517 1103164 254 273 3702 3721CTCACTGGCCCGGGTCTCCT 133  2648 1103180 485 504 3933 3952CTGTGCCAGATTGTCCCTCT  16* 2649 1103196 714 733 5600 5619CCACATGGACCTGCTGTCGG 121  2650 1103212 1046 1065 7618 7637TTCCTCCAGCCGCGCCAGCG 135  2651 1103228 1324 1343 11613 11632TGCTTGGACTCCTTAATGAC 97 2652 1103244 1436 1455 11725 11744GGGAAATGTGCCAGCAGAGG 39 2653 1103260 1601 1620 11890 11909CCCTCCAGACTGCCCCTTGG 95 2654 1103276 1726 1745 12015 12034CCATCTCTGGCAACAGTTTC 71 2655 1103292 1856 1875 12145 12164AAGACAAAACAAGCCTCTGG 88 2656 1103308 1882 1901 12171 12190GGGATATCCCACCTCATAAA 88 2657 1103324 2031 2050 12320 12339CCACCCATCTTAGACTGATC 100  2658 1103340 2198 2217 12487 12506ACACAACAGATCCCCCAAGT 90 2659 1103356 2410 2429 12699 12718TAATTCCCACAATCCAGAGG 115  2660 1103372 2459 2478 12748 12767TCCCCTTTCTCTCCTGTTTC 101  2661 1103388 2508 2527 12797 12816AGAGGCCTTTAGAAATGGGA 102  2662 1103404 2662 2681 12951 12970AAGAAGGGACCGCAAGAGGC 83 2663 1103420 2715 2734 13004 13023CAATTGTAAAATAGGGCACT 115  2664 1103436 2835 2854 13124 13143CAGTCTGCTCAGTCAAAGCA 121  2665 1103468 N/A N/A 8766 8785ATCTTGTGACCTTGTGATTT 75 2666 1103484 N/A N/A 8895 8914CTCACTCCCTGTCAAGCTGG 98 2667 1103500 N/A N/A 9033 9052CAGTTTAATGTACAGTTACT 90 2668 1103516 N/A N/A 9215 9234GCGATGGAGCCTCAGGGATG 28 2669 1103532 N/A N/A 9254 9273GAGGGAAGAATCCTCTGAAC 86 2670 1103548 N/A N/A 9305 9324CATTAAGAGCAGGGAACATA 85 2671 1103564 N/A N/A 8492 8511GAAGGCCCCCAGGGAGAGCT 72 2672 1103580 N/A N/A 8553 8572CAAGCCCCACCTAGAAGTAC 107  2673 1103596 N/A N/A 3966 3985CCCTCCTCACTTCTGCCTCA  98* 2674 1103612 N/A N/A 4047 4066TTCGGCCCCTCCCTGAGACT 77 2675 1103628 N/A N/A 4227 4246CCGCTGCTCCTGCACTGCTT 123  2676 1103644 N/A N/A 4387 4406CCTGCTCTTTCCCTCACTTT 110  2677 1103660 N/A N/A 4485 4504TATTATGACCACCGCTTCAC 85 2678 1103676 N/A N/A 4604 4623CACACACGCACATGTCCTCC 106  2679 1103692 N/A N/A 4754 4773GGGCCTTAGACAGAGGACTT 97 2680 1103708 N/A N/A 4970 4989CCTCCACCTCCCTGACCTGT  91* 2681 1103724 N/A N/A 5302 5321CAGTTGCAATCTCTGTGTTG 127  2682 1103740 N/A N/A 5392 5411TTCCTGTCTCTACCTGCCAA 153  2683 1103756 N/A N/A 5475 5494CTTCTGCCTGCCCCTCGGCC 109  2684 1103772 N/A N/A 5558 5577GAACCTCCTGACCAGGGTGA 116  2685 1103788 N/A N/A 5840 5859CTCTGGTGAAAGTCAGTCAC 112  2686 1103804 N/A N/A 5911 5930GTAGCAGTAATAATAATGGG 93 2687 1103820 N/A N/A 5969 5988CATGAAAAGCACAGGGCTTA 94 2688 1103836 N/A N/A 6017 6036GCTCTCTCATCTGTCAAAGA 59 2689 1103852 N/A N/A 6197 6216AGACACCTCTCTGTGTCCTG 60 2690 1103868 N/A N/A 6255 6274GTGGTGTTCTCTACGGGCAC 95 2691 1103884 N/A N/A 6308 6327GCCCCCTCTACAGTGTCTTT 85 2692 1103900 N/A N/A 6405 6424TCTGTGCTTTTCTGCCTCCA 59 2693 1103916 N/A N/A 6493 6512CTCACCCTGCTCAGACACCA 105  2694 1103932 N/A N/A 6900 6919CGAGCGGAGGCCTGGGTGTT 59 2695 1103948 N/A N/A 7006 7025CTAGCACAACACCTGGTCAG 106  2696 1103964 N/A N/A 7195 7214AGGTCTGCAAACTAGGTGGG 93 2697 1103980 N/A N/A 7414 7433CCCGCCCTCGACCCAGGTCC 121  2698 1103996 N/A N/A 7802 7821GAGCTCTACCGTGAGGCAGC 101  2699 1104012 N/A N/A 7911 7930ATCTAGTGACTGCCTGCTAT 95 2700 1104028 N/A N/A 8031 8050AGTCCAATCTTGGCTGGGAA 80 2701 1104044 N/A N/A 8073 8092GAAGAAAGTTCCAAGGAGGC 94 2702 1104060 N/A N/A 8209 8228TAACTCCATCTCCTAGCTTT 89 2703 1104076 N/A N/A 9349 9368CCCGAGAGAGAAAAATATAA 99 2704 1104092 N/A N/A 9406 9425TCATTTCAGGGCCAATGCAA 63 2705 1104108 N/A N/A 9450 9469TATTCTAGTCCAGAAGAAGA 127  2706 1104124 N/A N/A 9504 9523AGCCTTTCTGATGCTGAATT 52 2707 1104140 N/A N/A 9528 9547TTACCTCTACTAGTCAGCCT 75 2708 1104156 N/A N/A 9555 9574CCTGTGACTATCTAGGATTT 52 2709 1104172 N/A N/A 9574 9593TTTATCTGTGCTTTAGTGAC 50 2710 1104188 N/A N/A 9651 9670CATGTGGCACATATTAGTGC 97 2711 1104204 N/A N/A 9704 9723TACTTCTCTAGGTGGGAGAG 83 2712 1104220 N/A N/A 10137 10156AGTAAGACTCCGTCGAAAGC 124  2713 1104236 N/A N/A 10574 10593CTTTCTGAAAACCCAGCACG 87 2714 1104252 N/A N/A 10731 10750ATGAGAACCTATGCAACCGA 68 2715 1104268 N/A N/A 10807 10826CTCATGGACTTTCAGGGCAT 84 2716 1104284 N/A N/A 11076 11095TTACATTCAGTTTCCTTGCT 108  2717 1104300 N/A N/A 11147 11166GGATCCCATCTAGTGGCTTT 94 2718 1104316 N/A N/A 11222 11241TAACTTTAATTCTCTTTCTC 135  2719 1104332 N/A N/A 11266 11285CCCCATCCCCAACTGTGTCT 80 2720 1104348 N/A N/A 11385 11404TAAGCTGCTGGAGTAAGATG 92 2721 1104364 N/A N/A 11484 11503TTACTTTTCTTGATAGTAAC 89 2722 1104380 N/A N/A 11573 11592GCACAGTGCAACAGTTAGGA 38 2723

TABLE 37 Reduction of GFAP RNA by 5-10-5 MOE gapmers with mixedPO/PS internucleoside linkages in U251 cells SEQ ID SEQ ID SEQ ID SEQ IDNO: 1 NO: 1 NO: 2 NO: 2 SEQ Compound Start Stop Start Stop GFAP ID No.Site Site Site Site Sequence (5′ to 3′) (% UTC) NO 1047582 3047 306613336 13355 GTCTTTATTTTTCCTCAGCG  6 517 1103165 261 280 3709 3728CTGCCCGCTCACTGGCCCGG 98 2724 1103181 505 524 3953 3972TGCCTCACAGTGGCCAGGTC  45* 2725 1103197 756 775 5642 5661CTTTCAGGGCTGCGGTGAGG 99 2726 1103213 1048 1067 7620 7639TCTTCCTCCAGCCGCGCCAG 53 2727 1103229 1325 1344 11614 11633CTGCTTGGACTCCTTAATGA 73 2728 1103245 1497 1516 11786 11805AAGCTGACCTAGGGACAGAG 106  2729 1103261 1602 1621 11891 11910CCCCTCCAGACTGCCCCTTG 73 2730 1103277 1727 1746 12016 12035TCCATCTCTGGCAACAGTTT 48 2731 1103293 1857 1876 12146 12165AAAGACAAAACAAGCCTCTG 71 2732 1103309 1883 1902 12172 12191AGGGATATCCCACCTCATAA 159  2733 1103325 2057 2076 12346 12365TGACTGCCCCAGGTGGCAGG 116  2734 1103341 2199 2218 12488 12507TACACAACAGATCCCCCAAG 86 2735 1103357 2411 2430 12700 12719TTAATTCCCACAATCCAGAG 104  2736 1103373 2460 2479 12749 12768ATCCCCTTTCTCTCCTGTTT 76 2737 1103389 2509 2528 12798 12817AAGAGGCCTTTAGAAATGGG 71 2738 1103405 2663 2682 12952 12971TAAGAAGGGACCGCAAGAGG 81 2739 1103421 2716 2735 13005 13024ACAATTGTAAAATAGGGCAC 81 2740 1103437 2837 2856 13126 13145ACCAGTCTGCTCAGTCAAAG 90 2741 1103469 N/A N/A 8767 8786TATCTTGTGACCTTGTGATT 54 2742 1103485 N/A N/A 8896 8915GCTCACTCCCTGTCAAGCTG 76 2743 1103501 N/A N/A 9042 9061AAGCTCTGCCAGTTTAATGT 57 2744 1103517 N/A N/A 9216 9235AGCGATGGAGCCTCAGGGAT 83 2745 1103533 N/A N/A 9255 9274TGAGGGAAGAATCCTCTGAA 92 2746 1103549 N/A N/A 9306 9325ACATTAAGAGCAGGGAACAT 105  2747 1103565 N/A N/A 8499 8518GTGTCACGAAGGCCCCCAGG 72 2748 1103581 N/A N/A 8556 8575CTGCAAGCCCCACCTAGAAG 94 2749 1103597 N/A N/A 3989 4008TCACAAGGCCCCCCTTCCCC 98 2750 1103613 N/A N/A 4048 4067GTTCGGCCCCTCCCTGAGAC 90 2751 1103629 N/A N/A 4228 4247CCCGCTGCTCCTGCACTGCT 83 2752 1103645 N/A N/A 4389 4408TGCCTGCTCTTTCCCTCACT 88 2753 1103661 N/A N/A 4489 4508ATTTTATTATGACCACCGCT 97 2754 1103677 N/A N/A 4625 4644TCACTGTTGCACACACACAC 124  2755 1103693 N/A N/A 4775 4794GCAGGAGGATTAAGGGTTGG 76 2756 1103709 N/A N/A 5010 5029TGGGTGGCCATCAATCCTTT 97 2757 1103725 N/A N/A 5303 5322TCAGTTGCAATCTCTGTGTT 65 2758 1103741 N/A N/A 5394 5413ATTTCCTGTCTCTACCTGCC 95 2759 1103757 N/A N/A 5480 5499GTCCTCTTCTGCCTGCCCCT 90 2760 1103773 N/A N/A 5722 5741CAGGGCTACCTTGGAGCGGT 71 2761 1103789 N/A N/A 5848 5867TCTCACTTCTCTGGTGAAAG 82 2762 1103805 N/A N/A 5912 5931AGTAGCAGTAATAATAATGG 86 2763 1103821 N/A N/A 5973 5992AATCCATGAAAAGCACAGGG 88 2764 1103837 N/A N/A 6018 6037GGCTCTCTCATCTGTCAAAG 24 2765 1103853 N/A N/A 6198 6217CAGACACCTCTCTGTGTCCT 108  2766 1103869 N/A N/A 6256 6275TGTGGTGTTCTCTACGGGCA 62 2767 1103885 N/A N/A 6312 6331AAATGCCCCCTCTACAGTGT 110  2768 1103901 N/A N/A 6406 6425CTCTGTGCTTTTCTGCCTCC 67 2769 1103917 N/A N/A 6500 6519TCGGGCCCTCACCCTGCTCA 92 2770 1103933 N/A N/A 6902 6921GGCGAGCGGAGGCCTGGGTG 12 2771 1103949 N/A N/A 7016 7035ACCTCAGCACCTAGCACAAC 90 2772 1103965 N/A N/A 7197 7216TCAGGTCTGCAAACTAGGTG 72 2773 1103981 N/A N/A 7415 7434CCCCGCCCTCGACCCAGGTC 131  2774 1103997 N/A N/A 7804 7823ATGAGCTCTACCGTGAGGCA 88 2775 1104013 N/A N/A 7913 7932ACATCTAGTGACTGCCTGCT 94 2776 1104029 N/A N/A 8034 8053AGGAGTCCAATCTTGGCTGG 74 2777 1104045 N/A N/A 8074 8093TGAAGAAAGTTCCAAGGAGG 95 2778 1104061 N/A N/A 8215 8234AAAGTCTAACTCCATCTCCT 131  2779 1104077 N/A N/A 9358 9377CCCGCCCCGCCCGAGAGAGA 83 2780 1104093 N/A N/A 9408 9427AATCATTTCAGGGCCAATGC 24 2781 1104109 N/A N/A 9451 9470ATATTCTAGTCCAGAAGAAG 123  2782 1104125 N/A N/A 9505 9524TAGCCTTTCTGATGCTGAAT 57 2783 1104141 N/A N/A 9529 9548ATTACCTCTACTAGTCAGCC 59 2784 1104157 N/A N/A 9556 9575ACCTGTGACTATCTAGGATT 82 2785 1104173 N/A N/A 9575 9594ATTTATCTGTGCTTTAGTGA 41 2786 1104189 N/A N/A 9652 9671ACATGTGGCACATATTAGTG 98 2787 1104205 N/A N/A 9705 9724CTACTTCTCTAGGTGGGAGA 79 2788 1104221 N/A N/A 10138 10157GAGTAAGACTCCGTCGAAAG 89 2789 1104237 N/A N/A 10608 10627TCAGCCCCTCTGCAAGCCCT 85 2790 1104253 N/A N/A 10732 10751TATGAGAACCTATGCAACCG 72 2791 1104269 N/A N/A 10808 10827TCTCATGGACTTTCAGGGCA 112  2792 1104285 N/A N/A 11077 11096ATTACATTCAGTTTCCTTGC 103  2793 1104301 N/A N/A 11148 11167AGGATCCCATCTAGTGGCTT 91 2794 1104317 N/A N/A 11224 11243ACTAACTTTAATTCTCTTTC 96 2795 1104333 N/A N/A 11274 11293AAAGCCCTCCCCATCCCCAA 128  2796 1104349 N/A N/A 11387 11406GGTAAGCTGCTGGAGTAAGA 26 2797 1104365 N/A N/A 11485 11504CTTACTTTTCTTGATAGTAA 36 2798 1104381 N/A N/A 11605 11624CTCCTTAATGACCTGCAGGG 74 2799

TABLE 38 Reduction of GFAP RNA by 5-10-5 MOE gapmers withmixed PO/PS internucleoside linkages in U251 cells SEQ SEQ ID ID NO: 3NO: 3 SEQ Compound Start Stop GFAP ID Number Site SiteSequence (5′ to 3 ) (% UTC) NO 1103451 1220 1239 CCCTCGAATCTGCAGGTTGG 942800 1103452 1227 1246 TTTTGCCCCCTCGAATCTGC 106 2801 1103453 1228 1247CTTTTGCCCCCTCGAATCTG 84 2802 1103454 1229 1248 GCTTTTGCCCCCTCGAATCT 952803 1103455 1231 1250 GTGCTTTTGCCCCCTCGAAT 59 2804 1103456 1233 1252TGGTGCTTTTGCCCCCTCGA 92 2805 1103457 1235 1254 TTTGGTGCTTTTGCCCCCTC 602806 1103554 1813 1832 TTAATATTTAACATTAAGAG 77 2807 1047595 1222 1241CCCCCTCGAATCTGCAGGTT 109 284 1047596 1223 1242 GCCCCCTCGAATCTGCAGGT 107362 1047597 1225 1244 TTGCCCCCTCGAATCTGCAG 65 440 1047598 1230 1249TGCTTTTGCCCCCTCGAATC 115 518 1047599 1232 1251 GGTGCTTTTGCCCCCTCGAA 43596 1047600 1234 1253 TTGGTGCTTTTGCCCCCTCG 86 674

Example 3: Effect of Modified Oligonucleotides on Human GFAP RNA InVitro, Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in U251 cells. Cultured U251 cells at a density of10,000 cells per well were treated using free uptake with variousconcentrations of modified oligonucleotide as specified in the tablesbelow. After a treatment period of approximately 48 hours, total RNA wasisolated from the cells and GFAP RNA levels were measured byquantitative real-time RTPCR. Human GFAP primer probe set RTS37485 wasused to measure RNA levels, as described above. GFAP RNA levels wereadjusted according to total RNA content, as measured by RIBOGREEN®.Results are presented as percent of GFAP RNA, relative to untreatedcontrol cells (% control). Where possible, the half maximal inhibitoryconcentration (IC₅₀) of each modified oligonucleotide was calculatedusing a linear regression on a log/linear plot of the data in Excel. Insome cases, an IC₅₀ could not be reliably calculated and the data pointis marked as “NC”. Modified oligonucleotides marked with an asterisk (*)indicate that the modified oligonucleotide is complementary to theamplicon region of the primer probe set. Additional assays may be usedto measure the potency and efficacy of the modified oligonucleotidescomplementary to the amplicon region.

TABLE 39 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)IC₅₀ Compound No. 125 nM 500 nM 2000 nM 8000 nM (μM) 1047225 76 57 47 422.0 1047257 60 51 39 33 0.5 1047258 51 50 38 29 0.2 1047386 99 82 41 261.9 1047387 109 73 33 13 1.3 1047448 85 122 99 94 NC 1047466 107 82 7746 7.8 1047497 120 117 107 136 NC 1047512 116 112 97 87 NC 1047584 94 5423 10 0.8 1047608 125 133 79 52 NC 1047609 88 82 57 33 3.0 1047610 92 7543 19 1.5 1047913 98 72 49 25 1.9 1048203 120 95 57 31 3.3 1048267 99100 93 70 NC 1048296 99 91 73 72 NC 1048344 77 97 88 86 NC 1048361 92115 85 109 NC

TABLE 40 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)IC₅₀ Compound No. 125 nM 500 nM 2000 nM 8000 nM (μM) 1047325 81 86 76 67NC 1047357 74 63 47 22 1.1 1047372 145 143 125 99 NC 1047373 105 73 4321 1.7 1047374 150 122 84 46 7.3 1047388 108 116 62 32 4.2 1047500 100105 74 60 NC 1047515 93 95 81 65 NC 1047518 125 99 81 51 NC 1047579 92113 81 53 NC 1047580 96 88 58 41 4.3 1047581 73 60 32 17 0.7 1047582 7229 12 7 0.3 1047584 100 60 23 11 0.6 1047613 107 107 79 44 NC 1047662 9986 83 47 NC 1047707 100 126 91 75 NC 1048027 82 83 59 56 NC 1048350 10485 57 36 3.4

TABLE 41 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)IC₅₀ Compound No. 125 nM 500 nM 2000 nM 8000 nM (μM) 1047235 80 81 68 57NC 1047328 91 79 55 39 3.5 1047362 110 77 52 20 2.0 1047391 87 79 61 455.5 1047394 78 74 52 36 2.6 1047503 92 94 80 59 NC 1047522 81 81 63 50NC 1047583 97 54 19 12 0.8 1047584 81 50 19 10 0.6 1047585 85 73 38 181.2 1047586 75 64 41 23 1.1 1047587 92 66 41 19 1.3 1047599 92 94 83 78NC 1047601 103 89 68 50 NC 1047762 99 84 70 71 NC 1047955 95 104 86 92NC 1048018 85 84 74 51 NC 1048351 98 97 75 59 NC 1048355 114 114 90 61NC

TABLE 42 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)IC₅₀ Compound No. 125 nM 500 nM 2000 nM 8000 nM (μM) 1047429 80 58 32 150.8 1047444 80 98 83 73 NC 1047492 92 99 79 58 NC 1047573 80 74 59 444.7 1047584 87 53 20 8 0.7 1047588 69 49 24 14 0.5 1047589 101 79 47 242.0 1047590 70 55 35 24 0.7 1047591 92 67 34 21 1.2 1047734 86 75 69 60NC 1047990 75 75 58 40 4.1 1048055 103 96 61 42 5.0 1048151 94 76 23 161.1 1048182 85 76 56 35 2.9 1048195 105 104 87 87 NC 1048196 94 91 73 62NC 1048228 91 81 87 93 NC 1048341 80 71 61 46 6.1 1048372 94 88 84 59 NC

TABLE 43 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)IC₅₀ Compound No. 125 nM 500 nM 2000 nM 8000 nM (μM) 1047321 94 91 84 79NC 1047337 94 89 68 61 NC 1047368 86 82 79 81 NC 1047385 86 80 63 58 NC1047432 89 84 75 89 NC 1047465 82 67 62 49 7.3 1047469 104 95 73 58 NC1047513 106 100 96 80 NC 1047576 82 83 63 49 NC 1047584 88 63 31 14 1.01047625 86 80 73 55 NC 1047675 101 88 92 82 NC 1047688 99 100 96 100 NC1047705 100 92 78 68 NC 1047866 90 81 71 78 NC 1048125 103 91 86 84 NC1048187 100 88 82 74 NC 1048188 99 64 80 81 NC 1048201 80 65 41 23 1.2

TABLE 44 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)IC₅₀ Compound No. 125 nM 500 nM 2000 nM 8000 nM (μM) 1047298 87 83 74 75NC 1047474 102 93 94 87 NC 1047523 88 76 61 42 4.6 1047582 79 37 18 120.4 1047584 100 72 43 20 1.6 1047607 92 85 68 56 NC 1047717 106 110 9280 NC 1047746 108 110 106 110 NC 1047790 91 86 81 70 NC 1047799 104 10095 93 NC 1047840 92 88 75 58 NC 1047859 96 87 87 78 NC 1047886 84 79 6868 NC 1047907 97 90 80 71 NC 1048093 97 89 93 90 NC 1048144 101 93 99108 NC 1048207 97 82 76 74 NC 1048223 91 77 77 68 NC 1048350 104 80 5744 4.4

TABLE 45 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 25 11 7 5NC 1072814 62 45 22 14 0.7 1072815 86 57 42 28 2.3 1072818 78 54 33 161.5 1072834 33 22 13 8 NC 1072835 54 30 15 9 NC 1072855 64 44 27 16 0.81072862 59 40 20 9 0.6 1072863 83 54 35 20 1.7 1072868 80 50 23 9 1.21072872 21 13 9 7 NC 1072886 88 60 42 21 2.2 1072986 84 68 22 11 1.71073003 93 79 71 52 NC 1073034 97 80 30 30 2.9 1073035 78 80 41 27 2.81073063 53 29 15 7 NC 1073106 87 76 57 39 5.2 1073107 79 65 58 33 3.7

TABLE 46 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 36 15 8 6NC 1072813 69 58 36 17 1.4 1072824 85 63 38 23 2.2 1072849 66 32 21 140.6 1072853 116 98 70 24 5.1 1072856 90 57 32 14 1.8 1072857 66 45 32 171.0 1072861 68 45 28 16 1.0 1072864 87 62 43 22 2.3 1072973 101 74 40 132.5 1072980 39 45 27 16 NC 1073009 87 75 83 35 8.2 1073033 87 44 19 241.4 1073045 60 43 24 12 0.7 1073060 68 43 28 16 0.9 1073064 61 41 31 190.7 1073065 69 61 36 18 1.5 1073077 95 70 44 23 2.8 1073093 57 33 20 100.5

TABLE 47 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 70 30 9 50.6 1103198 91 86 82 73 NC 1103246 102 84 81 62 NC 1103278 89 96 72 51NC 1103279 86 69 43 30 2.9 1103359 82 75 55 39 5.0 1103470 86 88 73 55NC 1103471 68 63 45 36 2.6 1103502 104 93 75 51 NC 1103567 88 66 45 232.5 1104014 91 91 56 56 NC 1104078 92 100 103 92 NC 1104127 67 52 42 331.6 1104144 93 63 34 16 2.1 1104158 50 51 55 23 1.0 1104159 65 59 40 251.6 1104174 91 93 63 43 7.9 1104175 69 46 23 13 0.9 1104191 84 76 52 344.0

TABLE 48 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 66 30 116 0.6 1103202 79 79 54 43 6.0 1103217 77 38 16 5 0.9 1103218 92 74 50 333.7 1103330 84 79 56 51 NC 1103345 112 125 111 149 NC 1103377 115 134142 151 NC 1103409 82 93 70 45 NC 1103570 79 64 43 24 2.3 1103571 101 7461 34 4.6 1103872 93 79 28 21 2.4 1103970 88 78 75 74 NC 1104096 109 11265 47 9.2 1104131 87 79 54 44 6.1 1104145 99 89 65 39 6.7 1104161 102 9075 52 NC 1104178 72 67 57 53 NC 1104225 87 82 57 39 5.4 1104307 97 87 5732 4.7

TABLE 49 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 58 29 107 0.4 1103236 81 62 27 12 1.6 1103253 68 56 36 37 1.8 1103268 86 62 3730 2.4 1103285 91 79 56 29 4.0 1103300 100 98 83 81 NC 1103365 83 86 6442 8.2 1103428 87 73 50 26 3.2 1103491 86 70 70 55 NC 1103620 90 62 6573 NC 1104037 77 92 66 38 7.8 1104116 85 63 42 23 2.3 1104118 99 74 3820 2.6 1104132 107 79 53 35 4.5 1104133 70 67 43 29 2.3 1104165 91 86 5932 4.8 1104276 94 71 18 15 1.9 1104309 73 60 36 20 1.6 1104310 82 57 3013 1.6

TABLE 50 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 69 30 117 0.6 1103241 87 71 60 50 NC 1103257 75 83 47 26 3.1 1103258 81 70 44 232.4 1103272 94 91 52 26 4.0 1103369 88 78 59 38 5.4 1103370 83 88 59 448.0 1103400 95 101 87 76 NC 1103448 91 73 49 32 3.6 1103465 88 68 47 313.1 1103592 96 111 75 48 NC  1103722* 21 24 21 19 NC 1103816 108 118 9576 NC 1104072 111 83 63 17 3.8 1104122 84 73 65 48 NC 1104153 78 83 5550 NC 1104280 111 84 55 25 3.9 1104312 91 71 52 32 3.7 1104361 87 81 6542 7.4

TABLE 51 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 60 36 128 0.5 1103244 99 79 55 42 5.7 1103259 85 70 42 26 2.6 1103322 87 82 6646 9.5 1103371 86 97 64 38 7.3 1103516 104 123 90 69 NC 1103837 82 62 4026 2.2 1103899 94 87 68 48 NC 1103932 74 83 68 64 NC 1103933 79 63 11 91.3 1104093 90 61 33 24 2.1 1104123 91 114 81 61 NC 1104156 83 86 71 55NC 1104173 87 89 60 42 7.2 1104298 105 102 80 57 NC 1104299 101 109 8472 NC 1104349 104 71 57 35 4.3 1104365 95 75 63 40 5.8 1104380 95 83 6145 7.3

TABLE 52 Dose-dependent percent of human GFAP RNA compared to untreatedcontrol in U251 cells by modified oligonucleotides GFAP RNA (% control)Compound No. 370 nM 1111 nM 3333 nM 10000 nM IC50 (μM) 1047582 63 26 106 0.5 1103254 93 85 62 37 5.8 1103255 59 49 23 14 0.8 1103256 84 54 2210 1.4 1103366 105 79 58 58 NC 1103368 92 79 44 21 2.9 1103430 97 86 6042 6.3 1103446 84 79 51 36 4.3 1103447 88 79 49 32 3.7 1103478 90 95 6246 9.1 1103558 82 92 59 32 5.2 1103591 73 69 51 31 3.0 1104038 68 51 3136 1.4 1104039 80 68 43 36 3.1 1104151 80 65 46 28 2.6 1104152 94 71 4722 2.8 1104166 94 70 45 27 3.0 1104311 70 60 40 22 1.7 1104326 97 85 6452 NC

Example 4: Tolerability of Modified Oligonucleotides Complementary toHuman GFAP in Wild-Type Mice, 3 Hour Study

Modified oligonucleotides described above were tested in wild-typefemale C57/B16 mice to assess the tolerability of the oligonucleotides.Wild-type female C57/B16 mice each received a single ICV dose of 700 μgof modified oligonucleotide listed in the table below. Each treatmentgroup consisted of 4 mice. A group of 4 mice received PBS as a negativecontrol for each experiment (identified in separate tables below). At 3hours post-injection, mice were evaluated according to seven differentcriteria. The criteria are (1) the mouse was bright, alert, andresponsive; (2) the mouse was standing or hunched without stimuli; (3)the mouse showed any movement without stimuli; (4) the mousedemonstrated forward movement after it was lifted; (5) the mousedemonstrated any movement after it was lifted; (6) the mouse respondedto tail pinching; (7) regular breathing. For each of the 7 criteria, amouse was given a subscore of 0 if it met the criteria and 1 if it didnot (the functional observational battery score or FOB). After all 7criteria were evaluated, the scores were summed for each mouse andaveraged within each treatment group. The results are presented in thetables below.

TABLE 53 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1047198 3 1047258 3 1047328 7 1047362 0 1047373 0 1047386 41047387 0 1047388 2 1047391 5 1047580 0 1047582 5 1047583 2 1047584 11047585 0 1047586 0 1047587 0 1047588 0 1047589 1 1047590 2 1047591 41047610 5 1048203 5 1048267 6 1048350 4

TABLE 55 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1047211 2 1047223 0 1047225 1 1047298 0 1047306 0 1047316 21047352 4 1047353 2 1047402 2 1047432 0 1047522 0 1047523 2 1047532 21047579 2 1047598 4 1047602 5 1047662 4 1047679 2 1047711 2 1047811 01047884 3 1048201 1 1048204 0 1048227 1

TABLE 54 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1047257 5 1047357 7 1047374 5 1047394 6 1047429 0 1047444 21047448 0 1047492 2 1047497 2 1047500 1 1047518 5 1047573 1 1047581 11047599 1 1047601 6 1047608 2 1047609 5 1047913 1 1047990 2 1048027 21048151 4 1048182 0 1048296 3 1048361 0

TABLE 56 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1072813 0 1072814 1 1072818 4 1072834 4 1072835 0 1072849 51072855 4 1072856 6 1072857 4 1072861 6 1072862 4 1072863 3 1072868 71072872 5 1072886 1 1072980 1 1072986 6 1073033 5 1073045 6 1073060 61073063 6 1073064 6 1073065 6 1073093 6

TABLE 57 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1103217 5 1103368 3 1103370 3 1103371 3 1103465 6 1103502 51103516 7 1103567 5 1103837 4 1103899 2 1104093 5 1104118 6 1104133 61104145 2 1104152 5 1104158 3 1104166 5 1104168 4 1104175 6 1104307 41104309 5 1104310 6 1104311 5 1104349 7

TABLE 58 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1103202 5 1103218 7 1103253 7 1103255 7 1103256 7 1103257 61103258 7 1103259 6 1103268 1 1103279 6 1103285 7 1103411 7 1103471 61103570 6 1103722 1 1103872 4 1103933 7 1104038 7 1104116 1 1104127 31104144 3 1104159 5 1104276 1

Example 5: Design of MOE Gapmer Modified Oligonucleotides with MixedPO/PS Internucleoside Linkages Complementary to a Human GFAP NucleicAcid

Modified oligonucleotides complementary to human GFAP nucleic acid weredesigned. The modified oligonucleotides in the table below are 6-10-4MOE gapmers. The gapmers are 20 nucleosides in length and have a centralgap segment that consists of ten 2′-β-D-deoxynucleosides, a 5′ wingsegment that consists of six 2′-β-D-MOE nucleosides, and a 3′ wingsegment that consists of four 2′-β-D-MOE nucleosides. The sugar motif ofthe gapmers is (from 5′ to 3′): eeeeeeddddddddddeeee; wherein ‘d’represents a 2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a2′-β-D-MOE sugar moiety. The gapmers have an internucleoside linkagemotif of (from 5′ to 3′): sooooossssssssssoss; wherein each “s”represents a phosphorothioate internucleoside linkage and each “o”represents a phosphodiester internucleoside linkage. Each cytosinenucleoside is a 5-methylcytosine.

TABLE 59 6-10-4 MOE gapmers with mixed PO/PS internucleosidelinkages complementary to human GFAP SEQ ID SEQ ID SEQ ID SEQ ID No: 1No: 1 No: 2 No: 2 Compound Start Stop Start Stop SEQ ID No.SEQUENCE (5′ to 3′) Site Site Site Site No. 1166991 CAGCCTGGTTAGCCTTTCTGN/A N/A 9514 9533 2017 1166992 TCAGCCTGGTTAGCCTTTCT N/A N/A 9515 95342094 1166993 GTCAGCCTGGTTAGCCTTTC N/A N/A 9516 9535 2171 1166994AGTCAGCCTGGTTAGCCTTT N/A N/A 9517 9536 1484 1166996 CTAGTCAGCCTGGTTAGCCTN/A N/A 9519 9538 1561 1166997 ACTAGTCAGCCTGGTTAGCC N/A N/A 9520 95392325 1166998 CAGTATTACCTCTACTAGTC N/A N/A 9533 9552 20 1166999GCAGTATTACCTCTACTAGT N/A N/A 9534 9553 88 1167000 GGCAGTATTACCTCTACTAGN/A N/A 9535 9554 1637 1167001 TGGCAGTATTACCTCTACTA N/A N/A 9536 9555166 1167002 TTGGCAGTATTACCTCTACT N/A N/A 9537 9556 1865 1167003TTTGGCAGTATTACCTCTAC N/A N/A 9538 9557 1941 1167004 ATTTGGCAGTATTACCTCTAN/A N/A 9539 9558 2018 1167024 GTGTATTAGGATCCCATCTA N/A N/A 11155 111742028 1167025 TGTGTATTAGGATCCCATCT N/A N/A 11156 11175 2105 1167026GTGTGTATTAGGATCCCATC N/A N/A 11157 11176 1569 1167027AGTGTGTATTAGGATCCCAT N/A N/A 11158 11177 2182 1167050CTTTATTTTTCCTCAGCGAC 3045 3064 13334 13353 361 1167051TCTTTATTTTTCCTCAGCGA 3046 3065 13335 13354 439 1167053TGTCTTTATTTTTCCTCAGC 3048 3067 13337 13356 595 1167054TTGTCTTTATTTTTCCTCAG 3049 3068 13338 13357 673 1167055TTTGTCTTTATTTTTCCTCA 3050 3069 13339 13358 751 1167056ATTTGTCTTTATTTTTCCTC 3051 3070 13340 13359 829 1166975TTGTGATTTTCCCCGTCTTT N/A N/A 8755 8774 908 1166984 ACATTCACTAATATTTAACAN/A N/A 9323 9342 1022 1166985 CACATTCACTAATATTTAAC N/A N/A 9324 9343 211166986 TCACATTCACTAATATTTAA N/A N/A 9325 9344 2321 1166987GTCACATTCACTAATATTTA N/A N/A 9326 9345 1177 1166988 CGTCACATTCACTAATATTTN/A N/A 9327 9346 2398 1166989 ACGTCACATTCACTAATATT N/A N/A 9328 93472808 1166990 CACGTCACATTCACTAATAT N/A N/A 9329 9348 2809 1167057CATTTGTCTTTATTTTTCCT 3052 3071 13341 13360 907 1167059AGCATTTGTCTTTATTTTTC 3054 3073 13343 13362 1063 1167060CAGCATTTGTCTTTATTTTT 3055 3074 13344 13363 1140 1166982GCTTTTGAGATATCTTGTGA N/A N/A 8777 8796 51 1167007 TGACTATCTAGGATTTGGCAN/A N/A 9551 9570 2480 1167011 CCTGTGACTATCTAGGATTT N/A N/A 9555 95742709 1167012 TTTATCTGTGCTTTAGTGAC N/A N/A 9574 9593 2710 1167017TGCCATTTATCTGTGCTTTA N/A N/A 9579 9598 1639 1167018 CTGCCATTTATCTGTGCTTTN/A N/A 9580 9599 1867 1174062 GCTTTTGCCCCCTGTAGTGA N/A N/A 8732 87512291 1174063 GTGCTTTTGCCCCCTGTAGT N/A N/A 8734 8753 2368 1174065TGGTGCTTTTGCCCCCTGTA N/A N/A 8736 8755 2445 1174066 TTGGTGCTTTTGCCCCCTGTN/A N/A 8737 8756 59 1167033 GCACAGTTCCCAGATACTCC 1756 1775 12045 12064426 1167034 GGCACAGTTCCCAGATACTC 1757 1776 12046 12065 504 1167035AGGCACAGTTCCCAGATACT 1758 1777 12047 12066 1425 1167036AAGGCACAGTTCCCAGATAC 1759 1778 12048 12067 1502 1167037AAAGGCACAGTTCCCAGATA 1760 1779 12049 12068 582 1167040CTCAGTTTTCCTCCAGCAGC 1792 1811 12081 12100 115 1167046GCCAGTGTCTTCACTTTGCT 2750 2769 13039 13058 1973 1167058GCATTTGTCTTTATTTTTCC 3053 3072 13342 13361 985

Modified oligonucleotides in the table below are 5-10-5 MOE gapmers withmixed P0/PS internucleoside linkages. The gapmers are 20 nucleosides inlength, wherein the central gap segment consists of ten2′-β-D-deoxynucleosides and the 5′ and 3′ wing segments each consist offive 2′-β-D-MOE modified nucleosides. The sugar motif for the gapmers is(from 5′ to 3′): eeeeeddddddddddeeeee; wherein ‘d’ represents a2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-β-D-MOE sugarmoiety. The gapmers have an internucleoside linkage motif of (from 5′ to3′): soooossssssssssooss; wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage. Each cytosine nucleoside is a 5-methylcytosine.

TABLE 60 5-10-5 MOE gapmers with a mixed PO/PS internucleosidelinkages complementary to human GFAP SEQ ID SEQ ID SEQ ID SEQ ID No: 1No: 1 No: 2 No: 2 Compound Start Stop Start Stop SEQ ID No.SEQUENCE (5′ to 3′) Site Site Site Site No. 1166900 CAGCCTGGTTAGCCTTTCTGN/A N/A 9514 9533 2017 1166901 TCAGCCTGGTTAGCCTTTCT N/A N/A 9515 95342094 1166902 GTCAGCCTGGTTAGCCTTTC N/A N/A 9516 9535 2171 1166903AGTCAGCCTGGTTAGCCTTT N/A N/A 9517 9536 1484 1166904 TAGTCAGCCTGGTTAGCCTTN/A N/A 9518 9537 2248 1166905 CTAGTCAGCCTGGTTAGCCT N/A N/A 9519 95381561 1166906 ACTAGTCAGCCTGGTTAGCC N/A N/A 9520 9539 2325 1166907CAGTATTACCTCTACTAGTC N/A N/A 9533 9552 20 1166909 GGCAGTATTACCTCTACTAGN/A N/A 9535 9554 1637 1166910 TGGCAGTATTACCTCTACTA N/A N/A 9536 9555166 1166911 TTGGCAGTATTACCTCTACT N/A N/A 9537 9556 1865 1166912TTTGGCAGTATTACCTCTAC N/A N/A 9538 9557 1941 1166913 ATTTGGCAGTATTACCTCTAN/A N/A 9539 9558 2018 1166932 TGTATTAGGATCCCATCTAG N/A N/A 11154 111731951 1166933 GTGTATTAGGATCCCATCTA N/A N/A 11155 11174 2028 1166934TGTGTATTAGGATCCCATCT N/A N/A 11156 11175 2105 1166885CTTGTGATTTTCCCCGTCTT N/A N/A 8756 8775 986 1166886 CCTTGTGATTTTCCCCGTCTN/A N/A 8757 8776 1064 1166887 TTGAGATATCTTGTGACCTT N/A N/A 8773 87921510 1166888 TTTGAGATATCTTGTGACCT N/A N/A 8774 8793 1280 1166890CTTTTGAGATATCTTGTGAC N/A N/A 8776 8795 1434 1166893 ACATTCACTAATATTTAACAN/A N/A 9323 9342 1022 1166894 CACATTCACTAATATTTAAC N/A N/A 9324 9343 211166895 TCACATTCACTAATATTTAA N/A N/A 9325 9344 2321 1166896GTCACATTCACTAATATTTA N/A N/A 9326 9345 1177 1166897 CGTCACATTCACTAATATTTN/A N/A 9327 9346 2398 1166898 ACGTCACATTCACTAATATT N/A N/A 9328 93472808 1166899 CACGTCACATTCACTAATAT N/A N/A 9329 9348 2809 1166916TGACTATCTAGGATTTGGCA N/A N/A 9551 9570 2480 1166920 CCTGTGACTATCTAGGATTTN/A N/A 9555 9574 2709 1166926 TGCCATTTATCTGTGCTTTA N/A N/A 9579 95981639 1166927 CTGCCATTTATCTGTGCTTT N/A N/A 9580 9599 1867 1166928TCTGCCATTTATCTGTGCTT N/A N/A 9581 9600 400 1166929 CTCTGCCATTTATCTGTGCTN/A N/A 9582 9601 1943 1174056 GCTTTTGCCCCCTGTAGTGA N/A N/A 8732 87512291 1174058 GGTGCTTTTGCCCCCTGTAG N/A N/A 8735 8754 1227 1174059TGGTGCTTTTGCCCCCTGTA N/A N/A 8736 8755 2445 1174060 TTGGTGCTTTTGCCCCCTGTN/A N/A 8737 8756 59 1174061 TTTGGTGCTTTTGCCCCCTG N/A N/A 8738 8757 25221166940 ACAGTTCCCAGATACTCCGA 1754 1773 12043 12062 270 1166946AAAGGCACAGTTCCCAGATA 1760 1779 12049 12068 582 1166948TCAGTTTTCCTCCAGCAGCC 1791 1810 12080 12099 37 1166954CCAGTGTCTTCACTTTGCTC 2749 2768 13038 13057 825 1199982GGTCCTAAATATTCTAGTCC N/A N/A 9459 9478 2093 1199983 TGGTCCTAAATATTCTAGTCN/A N/A 9460 9479 2170 1199984 GTGGTCCTAAATATTCTAGT N/A N/A 9461 94802813

The modified oligonucleotides in the table below are 4-10-6 MOE gapmers.The gapmers are 20 nucleosides in length and have a central gap segmentthat consists of ten 2′-β-D-deoxynucleosides, a 5′ wing segment thatconsists of four 2′-β-D-MOE nucleosides, and a 3′ wing segment thatconsists of six 2′-β-D-MOE nucleosides. The sugar motif of the gapmersis (from 5′ to 3′): eeeeddddddddddeeeeee; wherein ‘d’ represents a2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-β-D-MOE sugarmoiety. The gapmers have an internucleoside linkage motif of (from 5′ to3′): sooossssssssssoooss; wherein “s” represents a phosphorothioateinternucleoside linkage and “o” represents a phosphodiesterinternucleoside linkage. Each cytosine nucleoside is a 5-methylcytosine.

TABLE 61 4-10-6 MOE gapmers with a mixed PO/PS internucleosidelinkages complementary to human GFAP SEQ ID SEQ ID SEQ ID SEQ ID No: 1No: 1 No: 2 No: 2 Compound Start Stop Start Stop SEQ ID No.SEQUENCE (5′ to 3′) Site Site Site Site No. 1166809 CAGCCTGGTTAGCCTTTCTGN/A N/A 9514 9533 2017 1166810 TCAGCCTGGTTAGCCTTTCT N/A N/A 9515 95342094 1166812 AGTCAGCCTGGTTAGCCTTT N/A N/A 9517 9536 1484 1166813TAGTCAGCCTGGTTAGCCTT N/A N/A 9518 9537 2248 1166814 CTAGTCAGCCTGGTTAGCCTN/A N/A 9519 9538 1561 1166815 ACTAGTCAGCCTGGTTAGCC N/A N/A 9520 95392325 1166816 CAGTATTACCTCTACTAGTC N/A N/A 9533 9552 20 1166817GCAGTATTACCTCTACTAGT N/A N/A 9534 9553 88 1166818 GGCAGTATTACCTCTACTAGN/A N/A 9535 9554 1637 1166819 TGGCAGTATTACCTCTACTA N/A N/A 9536 9555166 1166820 TTGGCAGTATTACCTCTACT N/A N/A 9537 9556 1865 1166821TTTGGCAGTATTACCTCTAC N/A N/A 9538 9557 1941 1166822 ATTTGGCAGTATTACCTCTAN/A N/A 9539 9558 2018 1166841 TGTATTAGGATCCCATCTAG N/A N/A 11154 111731951 1166842 GTGTATTAGGATCCCATCTA N/A N/A 11155 11174 2028 1166843TGTGTATTAGGATCCCATCT N/A N/A 11156 11175 2105 1166845AGTGTGTATTAGGATCCCAT N/A N/A 11158 11177 2182 1166846GAGTGTGTATTAGGATCCCA N/A N/A 11159 11178 2259 1166847AGAGTGTGTATTAGGATCCC N/A N/A 11160 11179 2336 1166869TCTTTATTTTTCCTCAGCGA 3046 3065 13335 13354 439 1166870GTCTTTATTTTTCCTCAGCG 3047 3066 13336 13355 517 1166871TGTCTTTATTTTTCCTCAGC 3048 3067 13337 13356 595 1166872TTGTCTTTATTTTTCCTCAG 3049 3068 13338 13357 673 1166873TTTGTCTTTATTTTTCCTCA 3050 3069 13339 13358 751 1166874ATTTGTCTTTATTTTTCCTC 3051 3070 13340 13359 829 1166793TTGTGATTTTCCCCGTCTTT N/A N/A 8755 8774 908 1166794 CTTGTGATTTTCCCCGTCTTN/A N/A 8756 8775 986 1166795 CCTTGTGATTTTCCCCGTCT N/A N/A 8757 87761064 1166798 TTTTGAGATATCTTGTGACC N/A N/A 8775 8794 1357 1166799CTTTTGAGATATCTTGTGAC N/A N/A 8776 8795 1434 1166800 GCTTTTGAGATATCTTGTGAN/A N/A 8777 8796 51 1166802 ACATTCACTAATATTTAACA N/A N/A 9323 9342 10221166803 CACATTCACTAATATTTAAC N/A N/A 9324 9343 21 1166804TCACATTCACTAATATTTAA N/A N/A 9325 9344 2321 1166805 GTCACATTCACTAATATTTAN/A N/A 9326 9345 1177 1166806 CGTCACATTCACTAATATTT N/A N/A 9327 93462398 1166807 ACGTCACATTCACTAATATT N/A N/A 9328 9347 2808 1166808CACGTCACATTCACTAATAT N/A N/A 9329 9348 2809 1166867 TTTATTTTTCCTCAGCGACT3044 3063 13333 13352 283 1166875 CATTTGTCTTTATTTTTCCT 3052 3071 1334113360 907 1166877 AGCATTTGTCTTTATTTTTC 3054 3073 13343 13362 10631166878 CAGCATTTGTCTTTATTTTT 3055 3074 13344 13363 1140 1166823ACTATCTAGGATTTGGCAGT N/A N/A 9549 9568 2326 1166826 GTGACTATCTAGGATTTGGCN/A N/A 9552 9571 1408 1166831 ATTTATCTGTGCTTTAGTGA N/A N/A 9575 95942786 1166835 TGCCATTTATCTGTGCTTTA N/A N/A 9579 9598 1639 1174050GCTTTTGCCCCCTGTAGTGA N/A N/A 8732 8751 2291 1174051 GTGCTTTTGCCCCCTGTAGTN/A N/A 8734 8753 2368 1174053 TGGTGCTTTTGCCCCCTGTA N/A N/A 8736 87552445 1174054 TTGGTGCTTTTGCCCCCTGT N/A N/A 8737 8756 59 1166852GGCACAGTTCCCAGATACTC 1757 1776 12046 12065 504

The modified oligonucleotides in the table below are 5-8-5 MOE gapmers.The gapmers are 20 nucleosides in length and have a central gap segmentthat consists of eight 2′-β-D-deoxynucleosides, a 5′ wing segment thatconsists of five 2′-β-D-MOE nucleosides, and a 3′ wing segment thatconsists of five 2′-β-D-MOE nucleosides. The sugar motif of the gapmersis (from 5′ to 3′): eeeeeddddddddeeeee; wherein ‘d’ represents a2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-β-D-MOE sugarmoiety. The gapmers have an internucleoside linkage motif of (from 5′ to3′): sooosssssssssooss; wherein each “s” represents a phosphorothioateinternucleoside linkage and each “o” represents a phosphodiesterinternucleoside linkage. Each cytosine nucleobase is a 5-methylcytosine.

TABLE 62 5-8-5 MOE gapmers with a mixed PO/PS internucleosidelinkages complementary to human GFAP SEQ ID SEQ ID SEQ ID SEQ ID No: 1No: 1 No: 2 No: 2 Compound Start Stop Start Stop SEQ ID No.SEQUENCE (5′ to 3′) Site Site Site Site No. 1166719 GCAGTATTACCTCTACTAN/A N/A 9536 9553 2816 1166720 GGCAGTATTACCTCTACT N/A N/A 9537 9554 28171166721 TGGCAGTATTACCTCTAC N/A N/A 9538 9555 2818 1166749TCAGCCTGGTTAGCCTTT N/A N/A 9517 9534 2819 1166750 GTCAGCCTGGTTAGCCTT N/AN/A 9518 9535 2820 1166751 AGTCAGCCTGGTTAGCCT N/A N/A 9519 9536 28211166761 GTGTATTAGGATCCCATC N/A N/A 11157 11174 2822 1166762TGTGTATTAGGATCCCAT N/A N/A 11158 11175 2823 1166763 GTGTGTATTAGGATCCCAN/A N/A 11159 11176 2824 1166775 TTATTTTTCCTCAGCGAC 3045 3062 1333413351 2825 1166776 TTTATTTTTCCTCAGCGA 3046 3063 13335 13352 2826 1166777CTTTATTTTTCCTCAGCG 3047 3064 13336 13353 2827 1166778 TCTTTATTTTTCCTCAGC3048 3065 13337 13354 2828 1166779 GTCTTTATTTTTCCTCAG 3049 3066 1333813355 2829 1166780 TGTCTTTATTTTTCCTCA 3050 3067 13339 13356 2830 1166781TTGTCTTTATTTTTCCTC 3051 3068 13340 13357 2831 1166782 TTTGTCTTTATTTTTCCT3052 3069 13341 13358 2832 1166783 ATTTGTCTTTATTTTTCC 3053 3070 1334213359 2833 1166738 GTGATTTTCCCCGTCTTT N/A N/A 8755 8772 2835 1166739TGTGATTTTCCCCGTCTT N/A N/A 8756 8773 2836 1166740 GAGATATCTTGTGACCTT N/AN/A 8773 8790 2837 1166742 TTGAGATATCTTGTGACC N/A N/A 8775 8792 28391166746 CACATTCACTAATATTTA N/A N/A 9326 9343 2840 1166747TCACATTCACTAATATTT N/A N/A 9327 9344 2841 1166748 GTCACATTCACTAATATT N/AN/A 9328 9345 2842 1166784 CATTTGTCTTTATTTTTC 3054 3071 13343 13360 28431166785 GCATTTGTCTTTATTTTT 3055 3072 13344 13361 2844 1166786AGCATTTGTCTTTATTTT 3056 3073 13345 13362 2845 1166787 CAGCATTTGTCTTTATTT3057 3074 13346 13363 2846 1174012 TTGTGATTTTCCCCGTCT N/A N/A 8757 87742850 1174013 TTTGAGATATCTTGTGAC N/A N/A 8776 8793 2851 1174015ATTCACTAATATTTAACA N/A N/A 9323 9340 2852 1174016 CATTCACTAATATTTAAC N/AN/A 9324 9341 2853 1174018 CGTCACATTCACTAATAT N/A N/A 9329 9346 28541174020 AGCCTGGTTAGCCTTTCT N/A N/A 9515 9532 2856 1174024AGTATTACCTCTACTAGT N/A N/A 9534 9551 2859 1174026 TTGGCAGTATTACCTCTA N/AN/A 9539 9556 2861 1174036 TATTAGGATCCCATCTAG N/A N/A 11154 11171 28621174037 GTATTAGGATCCCATCTA N/A N/A 11155 11172 2863 1166744GCTTTTGAGATATCTTGT N/A N/A 8779 8796 2866 1166757 CCATTTATCTGTGCTTTA N/AN/A 9579 9596 2873 1166758 TGCCATTTATCTGTGCTT N/A N/A 9581 9598 28741166759 CTGCCATTTATCTGTGCT N/A N/A 9582 9599 2875 1166760TCTGCCATTTATCTGTGC N/A N/A 9583 9600 2876 1174029 ACTATCTAGGATTTGGCA N/AN/A 9551 9568 2886 1174030 TGTGACTATCTAGGATTT N/A N/A 9555 9572 28871174031 TATCTGTGCTTTAGTGAC N/A N/A 9574 9591 2888 1174034GCCATTTATCTGTGCTTT N/A N/A 9580 9597 2891

Example 6: Tolerability of Modified Oligonucleotides Complementary toHuman GFAP in Wild-Type Mice, 3 Hour Study

Modified oligonucleotides described above were tested in wild-typefemale C57/B16 mice to assess the tolerability of the oligonucleotides.Wild-type female C57/B16 mice each received a single ICV dose of 700 μgof modified oligonucleotide listed in the table below. Each treatmentgroup consisted of 4 mice. A group of 4 mice received PBS as a negativecontrol for each experiment (identified in separate tables below). At 3hours post-injection, mice were evaluated according to seven differentcriteria. The criteria are (1) the mouse was bright, alert, andresponsive; (2) the mouse was standing or hunched without stimuli; (3)the mouse showed any movement without stimuli; (4) the mousedemonstrated forward movement after it was lifted; (5) the mousedemonstrated any movement after it was lifted; (6) the mouse respondedto tail pinching; (7) regular breathing. For each of the 7 criteria, amouse was given a subscore of 0 if it met the criteria and 1 if it didnot (the functional observational battery score or FOB). After all 7criteria were evaluated, the scores were summed for each mouse andaveraged within each treatment group. The results are presented in thetables below.

TABLE 63 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166719 1 1166720 0 1166721 0 1166749 3 1166750 1 1166751 21166761 4 1166762 4 1166763 3 1166775 0 1166776 1 1166777 4 1166778 11166779 4 1166780 1 1166781 0 1166782 0 1166783 0 1166809 4 1166810 41166812 3 1166813 3

TABLE 64 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166814 4 1166815 2 1166816 1 1166817 1 1166818 3 1166819 11166820 1 1166821 1 1166822 3 1166841 3 1166842 3 1166843 3 1166845 51166846 4 1166847 5 1166869 1 1166870 3 1166871 2 1166872 2 1166873 11166874 0 1166900 5

TABLE 65 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166901 4 1166902 5 1166903 4 1166904 4 1166905 2 1166906 21166907 0 1166909 3 1166910 1 1166911 0 1166912 1 1166913 2 1166932 41166933 2 1166934 3 1166991 4 1166992 4 1166993 4 1166994 3

TABLE 66 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166996 4 1166997 4 1166998 0 1166999 0 1167000 3 1167001 01167002 0 1167003 0 1167004 0 1167024 4 1167025 3 1167026 4 1167027 41167050 1 1167051 0 1167053 1 1167054 4 1167055 0 1167056 0

TABLE 67 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166738 0 1166739 0 1166740 4 1166742 3 1166746 0 1166747 01166748 0 1166784 0 1166785 1 1166786 3 1166787 3 1166793 0 1166794 01166795 1 1166797 4 1166798 3

TABLE 68 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166799 1 1166802 0 1166803 0 1166804 0 1166805 0 1166806 01166807 0 1166808 0 1166867 0 1166877 1 1166878 2 1166885 3 1166886 01166887 3 1166888 3 1166890 0 1166893 0

TABLE 69 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166894 0 1166895 0 1166896 0 1166897 0 1166898 0 1166899 01166984 0 1166985 0 1166986 0 1166987 0 1166988 0 1166990 0

TABLE 70 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1167057 0 1167059 1 1167060 1 1174012 2 1174013 3 1174015 01174016 0 1174018 0 1174020 4 1174024 3 1174026 0 1174036 3 1174037 2

TABLE 71 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166744 3 1166757 4 1166758 3 1166759 1 1166760 3 1166800 11166823 2 1166826 3 1166831 2 1166875 0 1166975 4 1166989 0

TABLE 72 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166835 0 1166916 3 1166920 2 1166926 1 1166927 2 1166928 11166929 0 1166982 2

TABLE 73 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1167007 2 1167011 1 1167012 3 1167017 2 1167018 1 1174029 4

TABLE 74 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1174030 3 1174031 3 1174034 1 1174050 3 1174051 3 1174053 11174054 1 1174056 4 1174058 3 1174059 0 1174060 0 1174061 2 1174062 31174063 3 1174065 1 1174066 1

TABLE 75 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166852 1

TABLE 76 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166940 2 1166946 2 1166948 2

TABLE 77 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1166954 0 1167033 0 1167034 1 1167035 1 1167036 2 1167037 21167040 2

TABLE 78 Tolerability scores in mice at 700 μg dose Compound No. 3 hr.FOB PBS 0 1167046 1 1167058 1

TABLE 79 Tolerability scores in mice at 700 μg dose 3 hr. Compound No.FOB PBS 0 1199982 1 1199983 1 1199984 1

Example 7: Activity of Modified Oligonucleotides Complementary to HumanGFAP in Transgenic Mice

Modified oligonucleotides described above were tested in human GFAPtransgenic mouse model. Transgenic mice (line 73.7) was previouslydescribed in Messing A., et al., Fatal encephalopathy with astrocyteinclusions in GFAP transgenic mice, Am J Pathos. 1998, 152(2):391-398.

Treatment

The GFAP transgenic mice were divided into groups of 3-4 mice each. Eachmouse received a single ICV bolus of 300 μg of modified oligonucleotide.A group of 3-4 mice received PBS as a negative control.

RNA Analysis

One week post treatment, mice were sacrificed and RNA was extracted fromcortical brain tissue, and/or spinal cord for RTPCR analysis to measureamount of GFAP RNA using primer probe set RTS38621 ((forward sequenceAGTTGCAGTCCTTGACCTG, designated herein as SEQ ID NO: 14; reversesequence CAGCGCCTCCTGATAACTG, designated herein as SEQ ID NO: 15; probesequence ACGAGTCCCTGGAGAGGCAGATG, designated herein as SEQ ID NO: 16),which is a human specific primer-probe set that recognizes all isoformsof GFAP. Results are presented as percent human GFAP RNA relative to PBScontrol, normalized to mouse peptidylprolyl isomerase A (PPIA), alsoknown as cyclophilin A. Mouse PPIA was amplified using primer probe setm_cyclo24 (forward sequence TCGCCGCTTGCTGCA, designated herein as SEQ IDNO: 17; reverse sequence ATCGGCCGTGATGTCGA, designated herein as SEQ IDNO: 18; probe sequence CCATGGTCAACCCCACCGTGTTC, designated herein as SEQID NO: 19). In some cases, RTPCR value is not defined for a certainsample, and is labeled N.D. (Not Defined).

As shown in the table below, treatment with modified oligonucleotidesresulted in reduction of GFAP RNA in comparison to the PBS control.

TABLE 80 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100 100 1047198 44 441047362 15 21 1047373 10 16 1047374 8 16 1047386 14 19 1047394 48 431047429 N.D. N.D. 1047448 37 45 1047500 23 26 1047580 12 15 1047582 5 81047583 6 9 1047584 6 7 1047585 8 7 1047586 12 11 1047587 13 13 10475899 13 1047590 16 16 1047591 14 17 1047610 33 31 1048203 26 27 1048350 2824

TABLE 81 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100  100  1047211 71 661047225 65 61 1047298 34 27 1047316 29 24 1047402 30 25 1047432 28 291047518 49 34 1047522 23 19 1047532 40 30 1047579 27 17 1047581 11 101047598 77 50 1047599 21 15 1047601 47 30 1047602 N.D. N.D. 1047609 2718 1047662 34 27 1047679 58 54

TABLE 82 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100 100 1047711 45 381047884 57 62 1047913 75 66 1048027 64 53 1048151 69 64 1048182 14 101048201 6 7 1048204 21 18 1048227 42 28 1072855 N.D. N.D. 1072856 N.D.N.D. 1072857 28 14 1072861 28 23 1072862 53 33 1072863 51 28 1072886 4226 1072980 40 25 1072986 98 88 1073033 100 80 1073045 36 24 1073060 1916 1073063 19 15 1073064 29 20 1073065 16 15 1073093 11 8

TABLE 83 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100 100 1103217 56 431103465 31 19 1103502 47 31 1103516 42 27 1103567 23 18 1104093 13 121104118 N.D. N.D. 1104145 8 6 1104152 27 17 1104166 17 13 1104175 15 8

TABLE 84 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100  100  1072813 15 181072814 15 19 1103202 34 36 1103218 48 33 1103253 56 36 1103255 N.D.N.D. 1103256 35 41 1103257 46 43 1103258 N.D. N.D. 1103259 82 71 1103268N.D. N.D. 1104307 13 14 1104309 10 11 1104311 11 18 1104349 N.D. N.D.

TABLE 85 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100 100 1103279 20 141103285 16 9 1103411 65 32 1103471 32 39 1103570 6 4 1103872 42 181104116 6 3 1104127 15 10 1104144 17 8 1104159 17 10 1104276 53 411166719 41 31 1166720 12 8 1166721 7 8 1166749 13 11 1166750 10 71166751 12 9 1166761 12 5

TABLE 86 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166762 18 1166763 13 1166775 211166776 30 1166777 13 1166778 6 1166779 7 1166780 9 1166781 7 1166782 121166783 17 1166809 13 1166810 16 1166812 23 1166813 21 1166814 291166815 42 1166816 31 1166817 37 1166818 14 1166819 9 1166820 12 116682115 1166822 20 1166841 47

TABLE 87 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. SPINAL CORD CORTEX PBS 100 100 1166842 19 141166843 37 21 1166845 17 19 1166846 12 18 1166847 13 19 1166869 10 111166870 6 7 1166871 5 7 1166872 10 9 1166873 14 14 1166874 9 10 116690014 20 1166901 11 13 1166902 12 15 1166903 17 19 1166904 26 30 1166905 2428 1166906 50 56 1166907 28 32

TABLE 88 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166909 30 1166910 8 1166911 71166912 15 1166913 12 1166932 41 1166933 13 1166934 18 1166991 141166992 11 1166993 9 1166994 10 1166996 21 1166997 30 1166998 13 116699911 1167000 15

TABLE 89 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1167001 9 1167002 7 1167003 81167004 13 1167024 18 1167025 11 1167026 14 1167027 13 1167050 111167051 10 1167053 7 1167054 5 1167055 6 1167056 6

TABLE 90 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166738 27 1166739 27 1166740 261166742 43 1166746 94 1166747 23 1166748 20 1166784 30 1166785 131166786 19 1166787 19 1166793 23 1166794 32 1166795 51 1166798 251166799 33 1166802 38 1166803 31 1166804 29 1166805 7 1166806 23 116680719 1166808 14 1166867 13 1166877 7 1166878 13 1166885 20 1166886 25

TABLE 91 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166887 12 1166888 16 1166890 381166893 39 1166894 20 1166895 11 1166896 8 1166897 6 1166898 8 116689911 1166984 24 1166985 9 1166986 13 1166987 7 1166988 5 1166990 9 11670577 1167059 5 1167060 8 1174012 9 1174013 33 1174015 107

TABLE 92 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166744 14 1166757 7 1166758 341166759 9 1166760 16 1166800 8 1166823 7 1166826 6 1166831 16 1166835 471166875 7 1166916 14 1166920 18 1166975 6 1166989 4 1174016 66 117401811 1174020 12 1174024 52 1174026 7 1174036 43 1174037 24

TABLE 93 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166926 14 1166927 15 1166928 131166929 15 1166982 11 1167007 44 1167011 21 1167012 23 1167017 8 116701811 1174029 32 1174030 75 1174031 35 1174034 17 1174050 10 1174051 151174053 22 1174054 40 1174056 7 1174058 14 1174059 14 1174060 19 117406117 1174062 12 1174063 12

TABLE 94 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1166852 15 1166940 16 1166946 341166948 41 1166954 16 1167033 9 1167034 11 1167035 15 1167036 19 116703725 1167040 15

TABLE 95 Reduction of human GFAP RNA in transgenic mice GFAP RNA (%control) Compound No. CORTEX PBS 100 1167046 14 1167058 5 1174065 101174066 13 1199982 4 1199983 13 1199984 16

Example 8: Tolerability of Modified Oligonucleotides Complementary toHuman GFAP in Rats, 3 mg Dose

Modified oligonucleotides described above were tested in rats to assessthe tolerability of the oligonucleotides. Sprague Dawley rats eachreceived a single intrathecal (IT) dose of 3 mg of oligonucleotidelisted in the table below. Each treatment group consisted of 2-4 rats. Agroup of 2-4 rats received PBS as a negative control. At 3 hourspost-injection, movement in 7 different parts of the body were evaluatedfor each rat. The 7 body parts are (1) the rat's tail; (2) the rat'sposterior posture; (3) the rat's hind limbs; (4) the rat's hind paws;(5) the rat's forepaws; (6) the rat's anterior posture; (7) the rat'shead. For each of the 7 different body parts, each rat was given asub-score of 0 if the body part was moving or 1 if the body part wasparalyzed (the functional observational battery score or FOB). Aftereach of the 7 body parts were evaluated, the sub-scores were summed foreach rat and then averaged for each group. For example, if a rat's tail,head, and all other evaluated body parts were moving 3 hours after the 3mg IT dose, it would get a summed score of 0. If another rat was notmoving its tail 3 hours after the 3 mg IT dose but all other evaluatedbody parts were moving, it would receive a score of 1. Results arepresented as the average score for each treatment group.

TABLE 96 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1047198 4 1047362 3 1047386 5 1047580 2 1047583 1 1047584 21047585 0 1047586 0 1047587 0 1047589 2 1047590 2 1047591 4 1048350 3

TABLE 97 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1047429 2 1047448 1 1047500 3 1047581 2 1047599 2 1047609 51047913 4 1048027 3 1048151 2

TABLE 98 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1047316 4 1047402 3 1047522 0 1047532 3 1047579 4 1048201 21048204 0

TABLE 99 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1047298 3 1072813 0 1072814 4 1072855 4 1072857 4 1104116 01104145 2 1104175 4 1166721 0 1166750 4 1166751 4 1166871 2

TABLE 100 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1166720 1 1166775 2 1166778 2 1166780 2 1166781 1 1166782 11166783 1 1166819 2 1166820 3 1166821 3 1166842 4 1166869 2 1166872 21166873 0 1166874 0

TABLE 101 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1166748 0 1166785 2 1166793 1 1166805 1 1166806 2 1166807 01166910 3 1166911 2 1166912 3 1166933 3 1166998 2 1167050 4 1167053 31167055 2 1167056 1

TABLE 102 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1166808 0 1166867 1 1166877 2 1166878 3 1166894 1 1166895 01166896 2 1166897 2 1166898 0 1166899 0 1166985 0 1166990 0

TABLE 103 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1166759 2 1166760 2 1166800 3 1166823 4 1166831 5 1166875 11166920 3 1166926 2 1166927 2 1166928 2 1166982 3 1166986 1 1166987 11166988 5 1166989 1 1167001 3

TABLE 104 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1167002 2 1167003 3 1167017 3 1167018 3 1167057 1 1167059 21167060 2 1174012 3 1174018 1 1174026 2 1174034 3 1174059 1 1174060 11174061 4

TABLE 105 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1166852 3 1166940 2 1166946 2 1166948 3 1166954 2

TABLE 106 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1167033 4 1167034 3 1167035 3 1167036 2 1167037 2 1167040 31167046 3 1167058 2

TABLE 107 Tolerability scores in rats at 3 mg dose FOB Compound No. 3 hrPBS 0 1174065 1 1174066 2 1199982 1 1199983 1 1199984 2

Example 9: Potency of Modified Oligonucleotides Complementary to HumanGFAP in Transgenic Mice

Modified oligonucleotides described above were tested in human GFAPtransgenic mouse model. Transgenic mice (line 73.7) was previouslydescribed in Messing A., et al., Fatal encephalopathy with astrocyteinclusions in GFAP transgenic mice, Am J Pathos. 1998, 152(2):391-398.

Treatment

The GFAP transgenic mice were divided into groups of 4 mice each. Eachmouse received a single ICV bolus of modified oligonucleotide at thedoses indicated in tables below. A group of 4-8 mice received PBS as anegative control.

RNA Analysis

Two weeks post treatment, mice were sacrificed, and RNA was extractedfrom the cortex, spinal cord, and brainstem for RTPCR analysis of RNAexpression of GFAP using primer probe set RTS38621, described in Example6 above. Results are presented as percent change of RNA, relative to PBScontrol, normalized to mouse PPIA. The half maximal effective dose(ED₅₀) of each modified oligonucleotide was calculated using GraphPadPrism 6 software (GraphPad Software, San Diego, Calif.). ED₅₀ valueswere calculated from dose and individual animal GFAP mRNA levels usingcustom equation Motulsky: Agonist vs response−Variable slope (fourparameters) Y=Bottom+(Top−Bottom)/(1+(10{circumflex over ( )} logEC50/X){circumflex over ( )}HillSlope), with the following constraints:bottom>lowest value in data set in order to compare across ASOs (3, 5,and 3 for cortex, spinal cord, and brainstem, respectively), top=100,HillSlope<−1 and >−2.

As shown in the table below, treatment with modified oligonucleotidesresulted in dose-responsive reduction of GFAP RNA in comparison to thePBS control.

TABLE 108 Reduction of human GFAP RNA (all isoforms) in transgenic miceGFAP RNA (% control) ED₅₀ Compound No. Dose (μg) CORTEX (μg) PBS N/A 100N/A 1104145 1 77 8 3 74 10 53 30 19 100 8

TABLE 109 Reduction of human GFAP RNA (all isoforms) in transgenic miceGFAP RNA (% control) ED₅₀ (μg) Compound SPINAL BRAIN SPINAL BRAIN No.Dose (μg) CORTEX CORD STEM CORTEX CORD STEM PBS N/A 100 100 100 N/A N/AN/A 1072813 3 93 101 99 34 24 21 10 68 71 69 30 70 38 37 100 18 28 21300 10 19 13 1166721 1 116 111 94 25 25 12 3 95 104 89 10 91 73 51 30 3947 28 100 13 18 11 300 7 10 6 1166874 1 81 102 78 27 11 7 3 73 83 75 1077 50 45 30 65 32 22 100 12 14 10 300 5 9 6 1166895 3 108 93 93 41 93 3810 82 94 80 30 46 74 50 100 40 40 32

TABLE 110 Reduction of human GFAP RNA (all isoforms) in transgenic miceGFAP RNA (% control) ED50 (μg) Compound SPINAL BRAIN SPINAL BRAIN No.Dose (μg) CORTEX CORD STEM CORTEX CORD STEM PBS N/A 100 100 100 N/A N/AN/A 1166895 300 16 45 27 N/A N/A N/A 3 103 77 77 1166926 10 86 70 57 4732 17 30 67 62 49 100 27 27 20 300 13 21 11 1166985 1 105 101 100 46 3614 3 96 90 78 10 75 68 53 30 79 58 40 100 17 35 15 300 8 23 10 1167056 1102 87 86 10 5 3 3 81 60 42 10 54 35 20 30 20 16 9 100 5 8 5 300 3 7 41167058 1 98 82 80 19 4 3 3 79 59 44 10 79 21 20

TABLE 111 Reduction of human GFAP RNA (all isoforms) in transgenic miceGFAP RNA (% control) ED50 (μg) Compound SPINAL BRAIN SPINAL BRAIN No.Dose (μg) CORTEX CORD STEM CORTEX CORD STEM PBS N/A 100 100 100 N/A N/AN/A 1166927 3 91 92 89 44 24 15 10 98 62 57 30 62 52 35 100 25 26 14 30014 20 12 1166954 3 106 87 85 58 60 26 10 79 80 68 30 70 65 49 100 36 4627 300 20 25 17 1166986 3 114 113 101 35 42 21 10 87 88 66 30 52 56 38100 23 30 20 300 10 20 14 1166998 3 77 73 61 23 13  6 10 90 63 42 30 3631 20 1167058 30 34 14 9 N/A N/A N/A 100 9 7 6 300 5 6 4

TABLE 112 Reduction of human GFAP RNA (all isoforms) in transgenic miceGFAP RNA (% control) RTS38621 ED50 (μg) Compound SPINAL BRAIN SPINALBRAIN No. Dose (μg) CORTEX CORD STEM CORTEX CORD STEM PBS 100 100 100N/A N/A N/A 1166998 100 11 18 17 N/A N/A N/A 300 7 12 9 N/A N/A N/A1199983 3 75 83 71 18 16 15 10 72 57 62 30 37 38 38 100 17 27 22 300 1222 17

Example 10: Effect of Modified Oligonucleotides on Human GFAP RNA InVitro, Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in U251 cells. Cultured U251 cells at a density of30,000 cells per well were treated with electroporation with variousconcentrations of modified oligonucleotide as specified in the tablesbelow. After a treatment period of approximately 24 hours, total RNA wasisolated from the cells and GFAP RNA levels were measured byquantitative real-time RTPCR. Human GFAP primer probe sets RTS38621(described herein above), RTS38624 (forward sequence AACCGGATCACCATTCCC,designated herein as SEQ ID NO:2892; reverse sequenceCCTTGTGATTTTCCCCGTCT, designated herein as SEQ ID NO: 1064; probesequence TGCTTTTGCCCCCTCGAATCTG, designated herein as SEQ ID NO: 2894),and RTS38622 (forward sequence AACCGGATCACCATTCCC, designated herein asSEQ ID NO: 2892; reverse sequence GTCTTCACCACGATGTTCCTC, designatedherein as SEQ ID NO: 2896; probe sequence CACCAAGTCTGTGTCAGAAGGCCA,designated herein as SEQ ID NO: 2897) were used to measure RNA levels.GFAP RNA levels were normalized to total GAPDH, as measured by humanprimer probe set RTS104 (forward sequence GAAGGTGAAGGTCGGAGTC,designated herein as SEQ ID NO: 2898; reverse sequenceGAAGATGGTGATGGGATTTC, designated herein as SEQ ID NO: 2899; probesequence CAAGCTTCCCGTTCTCAGCC, designated herein as SEQ ID NO: 2900).Results are presented as percent of GFAP RNA, relative to untreatedcontrol cells (% UTC). The half maximal inhibitory concentration (IC₅₀)of each modified oligonucleotide was calculated using a linearregression on a log/linear plot of the data in Excel.

TABLE 113 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS38621 IC₅₀ No.123 nM 370 nM 1100 nM 3300 nM 10000 nM (μM) 1072813 111 77 62 28 16 1.61166721 81 63 30 19 15 0.6 1166874 86 62 34 15 15 0.6 1166895 91 94 6840 21 2.5 1166954 69 52 35 14 12 0.4 1047582 54 26 13 6 5 0.1

TABLE 114 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS38622 IC₅₀ No.123 nM 370 nM 1100 nM 3300 nM 10000 nM (μM) 1072813 84 70 34 20 9 0.71166721 78 47 27 15 11 0.4 1166874 80 49 27 9 7 0.4 1166895 89 83 62 3517 1.8 1166954 68 50 27 9 5 0.3 1047582 51 24 10 3 1 0.1

TABLE 115 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS38624 IC₅₀ No.123 nM 370 nM 1100 nM 3300 nM 10000 nM (μM) 1072813 98 86 78 61 54 10.61166721 87 66 43 28 21 1.0 1166874 70 90 57 50 54 9.0 1166895 88 90 7152 34 4.0 1166954 77 69 67 53 50 6.9 1047582 68 56 47 41 43 1.2

TABLE 116 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS38621 IC₅₀ No.123 nM 370 nM 1100 nM 3300 nM 10000 nM (μM) 1166985 88 79 49 33 18 1.31166986 82 71 55 31 23 1.3 1166998 74 97 37 23 15 1.1 1167056 64 38 2312 10 0.2 1199983 70 53 37 17 10 0.4 1048182 69 53 30 15 10 0.4

TABLE 117 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS38622 IC₅₀ No.123 nM 370 nM 1100 nM 3300 nM 10000 nM (μM) 1166985 79 71 44 28 16 0.91166986 83 68 53 30 23 1.2 1166998 72 94 37 23 15 1.0 1167056 64 38 21 96 0.2 1199983 72 62 40 18 11 0.6 1048182 79 50 36 19 13 0.5

TABLE 118 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS38624 IC₅₀ No.123 nM 370 nM 1100 nM 3300 nM 10000 nM (μM) 1166985 80 75 54 44 25 1.81166986 86 73 65 43 31 2.5 1166998 76 111 53 32 17 1.8 1167056 80 72 6253 55 10.6 1199983 77 65 52 31 20 1.0 1048182 92 69 52 33 20 1.3

Example 11: Effect of Modified Oligonucleotides on Human GFAP RNA InVitro, Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in U251 cells. Cultured U251 cells at a density of20,000 cells per well were treated using free uptake with variousconcentrations of modified oligonucleotide as specified in the tablesbelow. After a treatment period of approximately 48 hours, total RNA wasisolated from the cells and GFAP RNA levels were measured byquantitative real-time RTPCR. Human GFAP primer probe sets RTS37485(described herein above) was used to measure RNA levels. GFAP RNA levelswere normalized to total GAPDH, as measured by human primer probe setRTS104 (described herein above). Results are presented as percent ofGFAP RNA, relative to untreated control cells (% UTC). The half maximalinhibitory concentration (IC₅₀) of each modified oligonucleotide wascalculated using a linear regression on a log/linear plot of the data inExcel.

TABLE 113 Dose-dependent reduction of human GFAP RNA in U251 cells bymodified oligonucleotides Compound GFAP RNA (% UTC) RTS37485 IC₅₀ No.250 nM 740 nM 2220 nM 6670 nM 20000 nM (μM) 1072813 105 102 80 59 34 101166721 92 83 64 49 32 7 1166874 81 78 55 38 22 4 1166895 83 89 85 78 73950 1166926 97 93 79 73 58 31 1166927 89 86 78 76 52 28 1166954 92 90 7767 58 29 1166985 104 108 104 94 80 43 1166986 94 92 95 84 82 10631166998 91 91 77 59 42 12 1167056 77 55 32 17 12 1 1167058 85 59 41 1815 1 1199983 94 88 73 62 49 17

The invention claimed is:
 1. A modified oligonucleotide according to thefollowing chemical structure:

or a salt thereof.
 2. A modified oligonucleotide according to thefollowing chemical structure:


3. The modified oligonucleotide of claim 1, which is the sodium salt orthe potassium salt.
 4. A pharmaceutical composition comprising themodified oligonucleotide of claim 1 and a pharmaceutically acceptablediluent.
 5. The pharmaceutical composition of claim 4, wherein thepharmaceutically acceptable diluent is artificial cerebrospinal fluid orphosphate-buffered saline (PBS).
 6. The pharmaceutical composition ofclaim 5, wherein the pharmaceutical composition consists of the modifiedoligonucleotide and artificial cerebrospinal fluid.
 7. An oligomericcompound comprising a modified oligonucleotide according to thefollowing chemical notation:^(m)C_(es)A_(eo)G_(eo)T_(eo)A_(eo)T_(eo)T_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds) ^(m)C_(ds)T_(ds)A_(ds)^(m)C_(ds)T_(ds)A_(eo)G_(es)T_(es) ^(m)C_(e) (SEQ ID NO: 20)wherein:A=an adenine nucleobase, ^(m)C=a 5-methylcytosine nucleobase, G=aguanine nucleobase, T=a thymine nucleobase, e=a 2′-β-D-MOE sugar moiety,d=a 2′-β-D-deoxyribosyl sugar moiety, s=a phosphorothioateinternucleoside linkage, and o=a phosphodiester internucleoside linkage.8. The oligomeric compound of claim 7, comprising the modifiedoligonucleotide covalently linked to a conjugate group.
 9. Apharmaceutical composition comprising the oligomeric compound of claim7, and a pharmaceutically acceptable diluent.
 10. The pharmaceuticalcomposition of claim 9, wherein the pharmaceutically acceptable diluentis artificial cerebrospinal fluid or PBS.
 11. The pharmaceuticalcomposition of claim 10, wherein the pharmaceutical composition consistsof the oligomeric compound and artificial cerebrospinal fluid.
 12. Amethod comprising administering to an individual the pharmaceuticalcomposition of claim
 4. 13. A method of treating Alexander disease adisease, comprising administering to an individual having or at risk ofhaving Alexander disease a therapeutically effective amount of thepharmaceutical composition according to claim 4, thereby treatingAlexander disease.
 14. The method of claim 13, wherein at least onesymptom or hallmark of Alexander disease is ameliorated.
 15. The methodof claim 14, wherein at least one symptom or hallmark is one or more ofmotor delays, cognitive delays, paroxysmal deterioration, seizures,vomiting, swallowing difficulties, ataxic gait, palatal myoclonus,autonomic dysfunction, or the presence of intra-astrocytic inclusionscalled Rosenthal fibers.
 16. The method of claim 13, wherein thepharmaceutical composition is administered to the central nervous systemor systemically.
 17. The method of claim 13, wherein the pharmaceuticalcomposition is administered to the central nervous system andsystemically.
 18. The pharmaceutical composition of claim 5, wherein thepharmaceutical composition consists essentially of the modifiedoligonucleotide and artificial cerebrospinal fluid.
 19. Thepharmaceutical composition of claim 10, wherein the pharmaceuticalcomposition consists essentially of the oligomeric compound andartificial cerebrospinal fluid.
 20. The pharmaceutical composition ofclaim 5, wherein the pharmaceutical composition consists essentially ofthe modified oligonucleotide and PBS.
 21. The pharmaceutical compositionof claim 10, wherein the pharmaceutical composition consists essentiallyof the oligomeric compound and PBS.
 22. A pharmaceutical compositioncomprising the modified oligonucleotide of claim 2 and apharmaceutically acceptable diluent.
 23. The pharmaceutical compositionof claim 22, wherein the pharmaceutically acceptable diluent isartificial cerebrospinal fluid or PBS.
 24. The pharmaceuticalcomposition of claim 23, wherein the pharmaceutical composition consistsessentially of the modified oligonucleotide and artificial cerebrospinalfluid.
 25. The pharmaceutical composition of claim 23, wherein thepharmaceutical composition consists essentially of the modifiedoligonucleotide and PBS.
 26. A pharmaceutical composition comprising themodified oligonucleotide of claim 3 and a pharmaceutically acceptablediluent.
 27. The pharmaceutical composition of claim 26, wherein thepharmaceutically acceptable diluent is artificial cerebrospinal fluid orPBS.
 28. The pharmaceutical composition of claim 27, wherein thepharmaceutical composition consists essentially of the modifiedoligonucleotide and artificial cerebrospinal fluid.
 29. Thepharmaceutical composition of claim 27, wherein the pharmaceuticalcomposition consists essentially of the modified oligonucleotide andPBS.
 30. A population of modified oligonucleotides of claim 1, whereinall of the phosphorothioate internucleoside linkages of the modifiedoligonucleotide are stereorandom.
 31. A population of modifiedoligonucleotides of claim 2, wherein all of the phosphorothioateinternucleoside linkages of the modified oligonucleotide arestereorandom.
 32. A population of modified oligonucleotides of claim 3,wherein all of the phosphorothioate internucleoside linkages of themodified oligonucleotide are stereorandom.
 33. A population ofoligomeric compounds of claim 7, wherein all of the phosphorothioateinternucleoside linkages of the modified oligonucleotide arestereorandom.
 34. A pharmaceutical composition comprising the populationof modified oligonucleotides of claim 30 and a pharmaceuticallyacceptable diluent.
 35. The pharmaceutical composition of claim 34,wherein the pharmaceutically acceptable diluent is artificialcerebrospinal fluid or PBS.
 36. A pharmaceutical composition comprisingthe population of modified oligonucleotides of claim 31 apharmaceutically acceptable diluent.
 37. The pharmaceutical compositionof claim 36, wherein the pharmaceutically acceptable diluent isartificial cerebrospinal fluid or PBS.
 38. A pharmaceutical compositioncomprising the population of modified oligonucleotides of claim 32 and apharmaceutically acceptable diluent.
 39. The pharmaceutical compositionof claim 38, wherein the pharmaceutically acceptable diluent isartificial cerebrospinal fluid or PBS.
 40. A pharmaceutical compositioncomprising the population of oligomeric compounds of claim 33 and apharmaceutically acceptable diluent.
 41. The pharmaceutical compositionof claim 40, wherein the pharmaceutically acceptable diluent isartificial cerebrospinal fluid or PBS.
 42. The method of claim 12,wherein the individual has or is at risk of having Alexander disease.